Interleukin-3 (IL-3) multiple mutation polypeptides

ABSTRACT

The present invention relates to recombinant human interleukin-3 (hIL-3) variant or mutant proteins (muteins). These hIL-3 muteins contain amino acid substitutions and may also have amino acid deletions at both the N- and C- termini. The invention also relates to pharmaceutical compositions containing the hIL-3 muteins and methods for using them. Additionally, the present invention relates to recombinant expression vectors comprising nucleotide sequences encoding the hIL-3 muteins, related microbial expression systems, and processes for making the hIL-3 muteins using the microbial expression systems.  
     Included in the present invention are deletion mutants of hIL-3 in which from 1 to 14 amino acids have been deleted from the N-terminus, and from 1 to 15 amino acids 119 to 133 have been deleted from the C-terminus, and which also contain amino acid substitutions in the polypeptide. These hIL-3 multiple mutation polypeptides may have biological activities similar to or better than hIL-3 and, in some cases, may also have an improved side effect profile.

[0001] This is a continuation-in-part of U.S. application Ser. No.07/981,044 filed Nov. 24, 1992 which is incorporated herein byreference.

FIELD OF THE INVENTION

[0002] The present invention relates to mutants or variants of humaninterleukin-3 (hIL-3) which contain multiple amino acid substitutionsand which may have portions of the native hIL-3 molecule deleted. ThesehIL-3 multiple mutation polypeptides retain one or more activities ofnative hIL-3 and may also show improved hematopoietic cell-stimulatingactivity and/or an improved activity profile which may include reductionof undesirable biological activities associated with native hIL-3.

BACKGROUND OF THE INVENTION

[0003] Colony stimulating factors (CSFs) which stimulate thedifferentiation and/or proliferation of bone marrow cells have generatedmuch interest because of their therapeutic potential for restoringdepressed levels of hematopoietic stem cell-derived cells. CSFs in bothhuman and murine systems have been identified and distinguishedaccording to their activities. For example, granulocyte-CSF (G-CSF) andmacrophage-CSF (M-CSF) stimulate the in vitro formation of neutrophilicgranulocyte and macrophage colonies, respectively while GM-CSF andinterleukin-3 (IL-3) have broader activities and stimulate the formationof both macrophage, neutrophilic and eosinophilic granulocyte colonies.IL-3 also stimulates the formation of mast, megakaryocyte and pure andmixed erythroid colonies.

[0004] Because of its ability to stimulate the proliferation of a numberof different cell types and to support the growth and proliferation ofprogenitor cells, IL-3 has potential for therapeutic use in restoringhematopoietic cells to normal amounts in those cases where the number ofcells has been reduced due to diseases or to therapeutic treatments suchas radiation and chemotherapy.

[0005] Interleukin-3 (IL-3) is a hematopoietic growth factor which hasthe property of being able to promote the survival, growth anddifferentiation of hematopoietic cells. Among the biological propertiesof IL-3 are the ability (a) to support the growth and differentiation ofprogenitor cells committed to all, or virtually all, blood celllineages; (b) to interact with early multipotential stem cells; (c) tosustain the growth of pluripotent precursor cells; (d) to stimulateproliferation of chronic myelogenous leukemia (CML) cells; (e) tostimulate proliferation of mast cells, eosinophils and basophils; (f) tostimulate DNA synthesis by human acute myelogenous leukemia (AML) cells;(g) to prime cells for production of leukotrienes and histamines; (h) toinduce leukocyte chemotaxis; and (i) to induce cell surface moleculesneeded for leukocyte adhesion.

[0006] Mature human interleukin-3 (hIL-3) consists of 133 amino acids.It has one disulfide bridge and two potential glycosylation sites (Yang,et al., CELL 47:3 (1986)).

[0007] Murine IL-3 (mIL-3) was first identified by Ihle, et al., J.IMMUNOL. 126:2184 (1981) as a factor which induced expression of a Tcell associated enzyme, 20_-hydroxysteroid dehydrogenase. The factor waspurified to homogeneity and shown to regulate the growth anddifferentiation of numerous subclasses of early hematopoietic andlymphoid progenitor cells.

[0008] In 1984, cDNA clones coding for murine IL-3 were isolated (Fung,et al., NATURE 307:233 (1984) and Yokota, et al., PROC. NATL. ACAD. SCI.USA 81:1070 (1984)). The murine DNA sequence coded for a polypeptide of166 amino acids including a putative signal peptide.

[0009] The gibbon IL-3 sequence was obtained using a gibbon cDNAexpression library. The gibbon IL-3 sequence was then used as a probeagainst a human genomic library to obtain a human IL-3 sequence.

[0010] Gibbon and human genomic DNA homologues of the murine IL-3sequence were disclosed by Yang, et al., CELL 47:3 (1986). The humansequence reported by Yang, et al. included a serine residue at position8 of the mature protein sequence. Following this finding, othersreported isolation of Pro⁸ hIL-3 cDNAs having proline at position 8 ofthe protein sequence. Thus it appears that there may be two allelicforms of hIL-3.

[0011] Dorssers, et al., GENE 55:115 (1987), found a clone from a humancDNA library which hybridized with mIL-3. This hybridization was theresult of the high degree of homology between the 3′ noncoding regionsof mIL-3 and hIL-3. This cDNA coded for an hIL-3 (Pro⁸) sequence.

[0012] U.S. Pat. No. 4,877,729 and U.S. Pat. No. 4,959,454 disclosehuman IL-3 and gibbon IL-3 cDNAs and the protein sequences for whichthey code. The hIL-3 disclosed has serine rather than proline atposition 8 in the protein sequence.

[0013] Clark-Lewis, et al., SCIENCE 231:134 (1986) performed afunctional analysis of murine IL-3 analogues synthesized with anautomated peptide synthesizer. The authors concluded that the stabletertiary structure of the complete molecule was required for fullactivity. A study on the role of the disulfide bridges showed thatreplacement of all four cysteines by alanine gave a molecule with{fraction (1/500)}th the activity as the native molecule. Replacement oftwo of the four Cys residues by Ala(Cys⁷⁹, Cys¹⁴⁰->Ala⁷⁹, Ala¹⁴⁰)resulted in an increased activity. The authors concluded that in murineIL-3 a single disulfide bridge is required between cysteines 17 and 80to get biological activity that approximates physiological levels andthat this structure probably stabilizes the tertiary structure of theprotein to give a conformation that is optimal for function.(Clark-Lewis, et al., PROC. NATL. ACAD. SCI. USA 85:7897 (1988)).

[0014] International Patent Application (PCT) WO 88/00598 disclosesgibbon- and human-like IL-3. The hIL-3 contains a Ser⁸->Pro⁸replacement. Suggestions are made to replace Cys by Ser, therebybreaking the disulfide bridge, and to replace one or more amino acids atthe glycosylation sites.

[0015] EP-A-0275598 (WO 88/04691) illustrates that Ala¹ can be deletedwhile retaining biological activity. Some mutant hIL-3 sequences areprovided, e.g., two double mutants, Ala¹->Asp¹, Trpl³->Arg¹³(pGB/IL-302) and Ala¹->Asp¹, Met³->Thr³ (pGB/IL-304) and one triplemutant Ala¹->Asp¹, Leu⁹->Pro⁹, Trp¹³->Arg¹³ (pGB/IL-303).

[0016] WO 88/05469 describes how deglycosylation mutants can be obtainedand suggests mutants of Arg⁵⁴Arg⁵⁵ and Arg¹⁰⁸Arg¹⁰⁹Lys¹¹⁰ might avoidproteolysis upon expression in Saccharomyces cerevisiae by KEX2protease. No mutated proteins are disclosed. Glycosylation and the KEX2protease activity are only important, in this context, upon expressionin yeast.

[0017] WO 88/06161 mentions various mutants which theoretically may beconformationally and antigenically neutral. The only actually performedmutations are Met²->Ile² and Ile¹³¹->Leu¹³¹. It is not disclosed whetherthe contemplated neutralities were obtained for these two mutations.

[0018] WO 91/00350 discloses nonglycosylated hIL-3 analog proteins, forexample, hIL-3 (Pro⁸Asp¹⁵Asp⁷⁰), Met³ rhul-3 (Pro⁸Asp¹⁵Asp⁷⁰); Thr⁴rhuL-3 (Pro⁸Asp¹⁵Asp⁷⁰)and Thr⁶ rhuIL-3 (Pro⁸Asp¹⁵Asp⁷⁰). It is saidthat these protein compositions do not exhibit certain adverse sideeffects associated with native hIL-3 such as urticaria resulting frominfiltration of mast cells and lymphocytes into the dermis. Thedisclosed analog hIL-3 proteins may have N termini at Met³, Thr⁴, orThr⁶.

[0019] WO 91/12874 discloses cysteine added variants (CAVs) of IL-3which have at least one Cys residue substituted for a naturallyoccurring amino acid residue.

SUMMARY OF THE INVENTION

[0020] The present invention relates to recombinant human interleukin-3(hIL-3) variant or mutant proteins (muteins). These hIL-3 muteinscontain amino acid substitutions and may also have amino acid deletionsat either/or both the N- and C- termini. Preferably, these mutantpolypeptides of the present invention contain four or more amino acidswhich differ from the amino acids found at the corresponding positionsin the native hIL-3 polypeptide. The invention also relates topharmaceutical compositions containing the hIL-3 muteins, DNA coding forthe muteins, and methods for using the muteins. Additionally, thepresent invention relates to recombinant expression vectors comprisingnucleotide sequences encoding the hIL-3 muteins, related microbialexpression systems, and processes for making the hIL-3 muteins using themicrobial expression systems.

[0021] The present invention includes mutants of hIL-3 in which from 1to 14 amino acids have been deleted from the N-terminus and/or from 1 to15 amino acids have been deleted from the C-terminus, and in whichmultiple amino acid substitutions have been made. Preferred muteins ofthe present invention are those in which amino acids 1 to 14 have beendeleted from the N-terminus, amino acids 126 to 133 have been deletedfrom the C-terminus, and which also contain from about four to abouttwenty-six amino acid substitutions in the polypeptide sequence. ThesehIL-3 multiple mutation polypeptides may have biological activitiessimilar to or better than hIL-3 and, in some cases, may also have animproved side effect profile, i.e., some muteins may have a bettertherapeutic index than native hIL-3. The present invention also providesmuteins which may function as IL-3 antagonists or as discrete antigenicfragments for the production of antibodies useful in immunoassay andimmunotherapy protocols. In addition to the use of the hIL-3 multiplemutation polypeptides of the present invention in vivo, it is envisionedthat in vitro uses would include the ability to stimulate bone marrowand blood cell activation and growth before infusion into patients.

[0022] Antagonists of hIL-3 would be particularly useful in blocking thegrowth of certain cancer cells like AML, CML and certain types of Blymphoid cancers. Other conditions where antagonists would be usefulinclude those in which certain blood cells are produced at abnormallyhigh numbers or are being activated by endogenous ligands. Antagonistswould effectively compete for ligands, presumably naturally occurringhemopoietins including and not limited to IL-3, GM-CSF and IL-5, whichmight trigger or augment the growth of cancer cells by virtue of theirability to bind to the IL-3 receptor complex while intrinsic activationproperties of the ligand are diminished. IL-3, GM-CSF and/or IL-5 alsoplay a role in certain asthmatic responses. An antagonist of the IL-3receptor may have the utility in this disease by blockingreceptor-mediated activation and recruitment of inflammatory cells.

BRIEF DESCRIPTION OF THE DRAWINGS

[0023]FIG. 1 is the human IL-3 gene for E. coli expression (pMON5873),encoding the polypeptide sequence of natural (wild type) human IL-3 [SEQID NO:128], plus an initiator methionine, as expressed in E. coli, withthe amino acids numbered from the N-terminus of the natural hIL-3.

[0024]FIG. 2: ClaI to NsiI Replacement Fragment. FIG. 2 shows thenucleotide sequence of the replacement fragment used between the ClaTand NsiI sites of the hIL-3 gene. The codon choice used in the fragmentcorresponds to that found in highly expressed E. coli genes (Gouy andGautier, 1982). Three new unique restriction sites, EcoRV, XhoI and PstIwere introduced for the purpose of inserting synthetic gene fragments.The portion of the coding sequence shown encodes hIL-3 amino acids20-70.

[0025]FIG. 3 shows the nucleotide and amino acid sequence of the gene inpMON5873 with the sequence extending from NcoI through HindIII. Thecodon choices used to encode amino acids 1-14 and 107-133 correspond tothat found in highly expressed E. coli genes.

[0026]FIG. 4 shows the construction of the plasmid vector pMON5846 whichencodes [Met-(1-133) hIL-3 (Arg¹²⁹)].

[0027]FIG. 5 shows the construction of the plasmid vector pMON5847 (ATCC68912) which encodes [Met-(1-133) hIL-3 (Arg¹²⁹)].

[0028]FIG. 6 shows the construction of plasmid vector pMON5853 whichencodes [Met-(15-133) hIL-3 (Arg¹²⁹)].

[0029]FIG. 7 shows the construction of the plasmid vector pMON5854 whichencodes [Met-(1-133) hIL-3 (Arg¹²⁹)].

[0030]FIG. 8 shows the DNA sequence and resulting amino acid sequence ofthe LamB signal peptide.

[0031]FIG. 9 shows the construction of the plasmid vector pMON5978 whichencodes Met-Ala-(15-125)hIL-3.

[0032]FIG. 10 shows the construction of the plasmid vector pMON5988which encodes Met-Ala(15-125)hIL-3.

[0033]FIG. 11 shows the construction of the plasmid vector pMON5887which encodes Met-(1-125)hIL-3.

[0034]FIG. 12 shows the construction of pMON6457 which encodes(15-125)hIL-3; it contains the araBAD promoter and the LamB signalpeptide fused to the variant hIL-3 amino acids 15-125.

[0035]FIG. 13 shows the construction of pMON6458; it contains the araBADpromoter and the LamB signal peptide fused to the variant hIL-3 aminoacids 15-125.

[0036]FIG. 14 shows the construction of pMON13359.

[0037]FIG. 15 shows the construction of pMON13352.

[0038]FIG. 16 shows the construction of pMON13360.

[0039]FIG. 17 shows the construction of pMON13363.

[0040]FIG. 18 shows the construction of pMON13364.

[0041]FIG. 19 shows the construction of pMON13365.

[0042]FIG. 20 shows the construction of pMON13287.

[0043]FIG. 21 shows the construction of pMON13288.

[0044]FIG. 22 shows the construction of pMON13289.

[0045]FIG. 23 shows the construction of pMON5723.

[0046]FIG. 24 shows the construction of pMON13438.

DETAILED DESCRIPTION OF THE INVENTION

[0047] The present invention relates to muteins of human interleukin-3(hIL-3) in which amino acid substitutions have been made at four or morepositions in amino acid sequence of the polypeptide and to muteins whichhave substantially the same structure and substantially the samebiological activity. Preferred muteins of the present invention are(15-125)hIL-3 deletion mutants which have deletions of amino acids 1 to14 at the N-terminus and 126 to 133 at the C-terminus and which alsohave four or more amino acid substitutions in the polypeptide andmuteins having substantially the same structure and substantially thesame biological activity. Among the preferred muteins are those havingtwenty-six amino acid substitutions. As used herein human interleukin-3corresponds to the amino acid sequence (1-133) as depicted in FIG. 1 and(15-125) hIL-3 corresponds to the 15 to 125 amino acid sequence of thehIL-3 polypeptide. Naturally occurring variants of hIL-3 polypeptideamino acids are also included in the present invention (for example, theallele in which proline rather than serine is at position 8 in the hIL-3polypeptide sequence) as are variant hIL-3 molecules which are modifiedpost-translationally (e.g. glycosylation).

[0048] The present invention also includes the DNA sequences which codefor the mutant polypeptides, DNA sequences which are substantiallysimilar and perform substantially the same function, and DNA sequenceswhich differ from the DNAs encoding the muteins of the invention onlydue to the degeneracy of the genetic code.

[0049] Included in the present invention are novel mutant humaninterleukin-3 polypeptides comprising a polypeptide having the aminoacid sequence of native human interleukin-3 wherein amino acids 126 to133 have been deleted from the C-terminus of the native humaninterleukin-3 polypeptide and amino acids 1 to 14 have been deleted fromthe N-terminus of the native human interleukin-3 polypeptide and, inaddition, polypeptides also have four or more amino acid substitutionsin the polypeptide sequence.

[0050] Also included in the present invention are the DNA sequencescoding for the muteins of the present invention; the oligonucleotideintermediates used to construct the mutant DNAs; and the polypeptidescoded for by these oligonucleotides. These polypeptides may be useful asantagonists or as antigenic fragments for the production of antibodiesuseful in immunoassay and immunotherapy protocols.

[0051] The mutant hIL-3 polypeptides of the present invention may alsohave methionine, alanine, or methionine-alanine residues inserted at theN-terminus.

[0052] The present invention includes human interleukin-3 mutantpolypeptide Formula I: Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr SerTrp Val Asn [SEQ ID NO:15] 1               5                   10                  15 Cys Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                 20                  25                  30 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa Xaa                 35                  40                  45 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                 50                  55                  60 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                 65                  70                  75 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                 80                  85                  90 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                 95                 100                 105 Xaa Phe XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                110                 115                 120 Xaa Xaa XaaGln Gln Thr Thr Leu Ser Leu Ala Ile Phe                125                 130

[0053] wherein

[0054] Xaa at position 17 is Ser, Lys, Gly, Asp, Met, Gln, or Arg;

[0055] Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

[0056] Xaa at position 19 is Met, Phe, Ile, Arg, Gly, Ala, or Cys;

[0057] Xaa at position 20 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;

[0058] Xaa at position 21 is Asp, Phe, Lys, Arg, Ala, Gly, Glu, Gln,Asn, Thr, Ser or Val;

[0059] Xaa at position 22 is Glu, Trp, Pro, Ser, Ala, His, Asp, Asn,Gln, Leu, Val or Gly;

[0060] Xaa at position 23 is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe,Leu, Ser, or Arg;

[0061] Xaa at position 24 is Ile, Gly, Val, Arg, Ser, Phe, or Leu;

[0062] Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

[0063] Xaa at position 26 is His, Thr, Phe, Gly, Arg, Ala, or Trp;

[0064] Xaa at position 27 is Leu, Gly, Arg, Thr, Ser, or Ala;

[0065] Xaa at position 28 is Lys, Arg, Leu, Gln, Gly, Pro, Val or Trp;

[0066] Xaa at position 29 is Gln, Asn, Leu, Pro, Arg, or Val;

[0067] Xaa at position 30 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, orLys;

[0068] Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

[0069] Xaa at position 32 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu;

[0070] Xaa at position 33 is Pro, Leu, Gln, Ala, Thr, or Glu;

[0071] Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Glu, Gln, Thr,Arg, Ala, Phe, Ile or Met;

[0072] Xaa at position 35 is Leu, Ala, Gly, Asn, Pro, Gln, or Val;

[0073] Xaa at position 36 is Asp, Leu, or Val;

[0074] Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile;

[0075] Xaa at position 38 is Asn, or Ala;

[0076] Xaa at position 40 is Leu, Trp, or Arg;

[0077] Xaa at position 41 is Asn, Cys, Arg, Leu, His, Met, or Pro;

[0078] Xaa at position 42 is Gly, Asp, Ser, Cys, Asn, Lys, Thr, Leu,Val, Glu, Phe, Tyr, Ile, Met or Ala;

[0079] Xaa at position 43 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys,Gln, Arg, Thr, Gly or Ser;

[0080] Xaa at position 44 is Asp, Ser, Leu, Arg, Lys, Thr, Met, Trp,Glu, Asn, Gln, Ala or Pro;

[0081] Xaa at position 45 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys,Trp, Asp, Asn, Arg, Ser, Ala, Ile, Glu or His;

[0082] Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Gln, Asn, Gln,Lys, His, Ala, Tyr, Ile, Val or Gly;

[0083] Xaa at position 47 is Ile, Gly, Val, Ser, Arg, Pro, or His;

[0084] Xaa at position 48 is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu,Lys, Thr, Ala, Met, Val or Asn;

[0085] Xaa at position 49 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;

[0086] Xaa at position 50 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser,Ala, Ile, Val, His, Phe, Met or Gln;

[0087] Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;

[0088] Xaa at position 52 is Asn, His, Arg, Leu, Gly, Ser, or Thr;

[0089] Xaa at position 53 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, orMet;

[0090] Xaa at position 54 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn,Lys, His, Ala or Leu;

[0091] Xaa at position 55 is Arg, Thr, Val, Ser, Leu, or Gly;

[0092] Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His,Thr, Ala, Tyr, Phe, Leu, Val or Lys;

[0093] Xaa at position 57 is Asn or Gly;

[0094] Xaa at position 58 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;

[0095] Xaa at position 59 is Glu Tyr, His, Leu, Pro, or Arg;

[0096] Xaa at position 60 is Ala, Ser, Pro, Tyr, Asn, or Thr;

[0097] Xaa at position 61 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser;

[0098] Xaa at position 62 is Asn His, Val, Arg, Pro, Thr, Asp, or Ile;

[0099] Xaa at position 63 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or Val;

[0100] Xaa at position 64 is Ala, Asn, Pro, Ser, or Lys;

[0101] Xaa at position 65 is Val, Thr, Pro, His, Leu, Phe, or Ser;

[0102] Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

[0103] Xaa at position 67 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, orHis;

[0104] Xaa at position 68 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or His;

[0105] Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly, orLeu;

[0106] Xaa at position 70 is Asn, Leu, Val, Trp, Pro, or Ala;

[0107] Xaa at position 71 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln,Trp, or Asn;

[0108] Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;

[0109] Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or Arg;

[0110] Xaa at position 74 is Ile, Met, Thr, Pro, Arg, Gly, Ala;

[0111] Xaa at position 75 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser,Gln, or Leu;

[0112] Xaa at position 76 is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly, orAsp;

[0113] Xaa at position 77 is Ile, Ser, Arg, Thr, or Leu;

[0114] Xaa at position 78 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg;

[0115] Xaa at position 79 is Lys, Thr, Asn, Met, Arg, Ile, Gly, or Asp;

[0116] Xaa at position 80 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or Arg;

[0117] Xaa at position 81 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys;

[0118] Xaa at position 82 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn,His, Thr, Ser, Ala, Tyr, Phe, Ile, Met or Val;

[0119] Xaa at position 83 is Pro, Ala, Thr, Trp, Arg, or Met;

[0120] Xaa at position 84 is Cys, Gln, Gly, Arg, Met, or Val;

[0121] Xaa at position 85 is Leu, Asn, Val, or Gln;

[0122] Xaa at position 86 is Pro, Cys, Arg, Ala, or Lys;

[0123] Xaa at position 87 is Leu, Ser, Trp, or Gly;

[0124] Xaa at position 88 is Ala, Lys, Arg, Val, or Trp;

[0125] Xaa at position 89 is Thr, Asp, Cys, Leu, Val, Glu, His, Asn, orSer;

[0126] Xaa at position 90 is Ala, Pro, Ser, Thr, Gly, Asp, Ile, or Met;

[0127] Xaa at position 91 is Ala, Pro, Ser, Thr, Phe, Leu, Asp, or His;

[0128] Xaa at position 92 is Pro, Phe, Arg, Ser, Lys, His, Ala, Gly, Ileor Leu;

[0129] Xaa at position 93 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;

[0130] Xaa at position 94 is Arg, Ile, Ser, Glu, Leu, Val, Gln, Lys,His, Ala, or Pro;

[0131] Xaa at position 95 is His, Gln, Pro, Arg, Val, Leu, Gly, Thr,Asn, Lys, Ser, Ala, Trp, Phe, Ile, or Tyr;

[0132] Xaa at position 96 is Pro, Lys, Tyr, Gly, Ile, or Thr;

[0133] Xaa at position 97 is Ile, Val, Lys, Ala, or Asn;

[0134] Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr, Glu,Gln, Ser, Phe, Met, Val, Lys, Arg, Tyr or Pro;

[0135] Xaa at position 99 is Ile, Leu, Arg, Asp, Val, Pro, Gln, Gly,Ser, Phe, or His;

[0136] Xaa at position 100 is Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln, orPro;

[0137] Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val, Tyr,Glu, Asn, Ser, Ala, Gly, Ile, Leu, or Gln;

[0138] Xaa at position 102 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;

[0139] Xaa at position 103 is Asp, or Ser;

[0140] Xaa at position 104 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu,Gln, Lys, Ala, Phe, or Gly;

[0141] Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,Leu, Lys, Ile, Asp, or His;

[0142] Xaa at position 106 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;

[0143] Xaa at position 108 is Arg, Lys, Asp, Leu, Thr, Ile, Gln, His,Ser, Ala or Pro;

[0144] Xaa at position 109 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly;

[0145] Xaa at position 110 is Lys, Ala, Asn, Thr, Leu, Arg, Gln, His,Glu, Ser, Ala, or Trp;

[0146] Xaa at position 111 is Leu, Ile, Arg, Asp, or Met;

[0147] Xaa at position 112 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe;

[0148] Xaa at position 113 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp,Lys, Leu, Ile, Val or Asn;

[0149] Xaa at position 114 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;

[0150] Xaa at position 115 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr,Trp, or Met;

[0151] Xaa at position 116 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu,Arg, Trp, Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile;

[0152] Xaa at position 117 is Thr, Ser, Asn, Ile, Trp, Lys, or Pro;

[0153] Xaa at position 118 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;

[0154] Xaa at position 119 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg;

[0155] Xaa at position 120 is Asn, Ala, Pro, Leu, His, Val, or Gln;

[0156] Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;

[0157] Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys;

[0158] Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

[0159] and which can additionally have Met- preceding the amino acid inposition 1; and wherein from 1 to 14 amino acids can be deleted from theN-terminus and/or from 1 to 15 amino acids can be deleted from theC-terminus; and wherein from 4 to 44 of the amino acids designated byXaa are different from the corresponding amino acids of native (1-133)human interleukin-3.

[0160] Included in the present invention are human interleukin-3 mutantpolypeptide of the Formula II: Ala Pro Met Thr Gln Thr Thr Ser Leu LysThr Ser Trp Val Asn [SEQ ID NO:16] 1           5               10                   15 Cys Xaa Xaa Xaa XaaXaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa Xaa            20               25                   30 Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa Xaa            35               40                   45 Xaa Xaa Leu Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa            50               55                   60 Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa            65               70                   75 Xaa Xaa Leu Xaa XaaXaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa Xaa            80               85                   90 Xaa Xaa Xaa Arg XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa             95             100            105 Xaa Phe Xaa Xaa Lys LeuXaa Phe Xaa Xaa Xaa Xaa Leu Xaa Xaa            110             115            120 Xaa Xaa Xaa Gln Gln ThrThr Leu Ser Leu Ala Ile Phe             125             130

[0161] wherein

[0162] Xaa at position 17 is Ser, Gly, Asp, Met, or Gln;

[0163] Xaa at position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

[0164] Xaa at position 19 is Met, Phe, Ile, Arg, or Ala;

[0165] Xaa at position 20 is Ile or Pro;

[0166] Xaa at position 21 is Asp or Glu;

[0167] Xaa at position 23 is Ile, Val, Ala, Leu, or Gly;

[0168] Xaa at position 24 is Ile, Val, Phe, or Leu;

[0169] Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

[0170] Xaa at position 26 is His, Phe, Gly, Arg, or Ala;

[0171] Xaa at position 28 is Lys, Leu, Gln, Gly, Pro, or Val;

[0172] Xaa at position 29 is Gln, Asn, Leu, Arg, or Val;

[0173] Xaa at position 30 is Pro, His, Thr, Gly, or Gln;

[0174] Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

[0175] Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

[0176] Xaa at position 33 is Pro, Leu, Gln, Ala, or Glu;

[0177] Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln,Glu, Ile, Phe, Thr or Met;

[0178] Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val;

[0179] Xaa at position 36 is Asp or Leu;

[0180] Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile;

[0181] Xaa at position 38 is Asn or Ala;

[0182] Xaa at position 41 is Asn, Cys, Arg, His, Met, or Pro;

[0183] Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu,Met, Tyr, Val or Arg;

[0184] Xaa at position 44 is Asp or Glu;

[0185] Xaa at position 45 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn,Glu, Ser, or Trp;

[0186] Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln,Glu, His, Ile, Lys, Tyr, Val or Gly;

[0187] Xaa at position 47 is Ile, Val, or His;

[0188] Xaa at position 49 is Met, Asn, or Asp;

[0189] Xaa at position 50 is Glu, Thr, Ala, Asn, Ser or Asp;

[0190] Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;

[0191] Xaa at position 52 is Asn or Gly;

[0192] Xaa at position 53 is Leu, Met, or Phe;

[0193] Xaa at position 54 is Arg, Ala, or Ser;

[0194] Xaa at position 55 is Arg, Thr, Val, Leu, or Gly;

[0195] Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Ala, Arg, Asn,Glu, His, Leu, Thr, Val or Lys;

[0196] Xaa at position 59 is Glu, Tyr, His, Leu, or Arg;

[0197] Xaa at position 60 is Ala, Ser, Asn, or Thr;

[0198] Xaa at position 61 is Phe or Ser;

[0199] Xaa at position 62 is Asn, Val, Pro, Thr, or Ile;

[0200] Xaa at position 63 is Arg, Tyr, Lys, Ser, His, or Val;

[0201] Xaa at position 64 is Ala or Asn;

[0202] Xaa at position 65 is Val, Thr, Leu, or Ser;

[0203] Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

[0204] Xaa at position 67 is Ser, Phe, Val, Gly, Asn, Ile, or His;

[0205] Xaa at position 68 is Leu, Val, Ile, Phe, or His;

[0206] Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

[0207] Xaa at position 70 is Asn or Pro;

[0208] Xaa at position 71 is Ala, Met, Pro, Arg, Glu, Thr, or Gln;

[0209] Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;

[0210] Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, Arg, orPro;

[0211] Xaa at position 74 is Ile or Met;

[0212] Xaa at position 75 is Glu, Gly, Asp, Ser, or Gln;

[0213] Xaa at position 76 is Ser, Val, Ala, Asn, Glu, Pro, Gly, or Asp;

[0214] Xaa at position 77 is Ile, Ser, or Leu;

[0215] Xaa at position 79 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, orAsp;

[0216] Xaa at position 80 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;

[0217] Xaa at position 81 is Leu, or Val;

[0218] Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu,His, Met, Phe, Ser, Thr, Tyr or Val;

[0219] Xaa at position 83 is Pro, Ala, Thr, Trp, or Met;

[0220] Xaa at position 85 is Leu or Val;

[0221] Xaa at position 87 is Leu or Ser;

[0222] Xaa at position 88 is Ala, Arg, or Trp;

[0223] Xaa at position 89 is Thr, Asp, Glu, His, Asn, or Ser;

[0224] Xaa at position 90 is Ala, Asp, or Met;

[0225] Xaa at position 91 is Ala, Pro, Ser, Thr, Phe, Leu, or Asp;

[0226] Xaa at position 92 is Pro or Ser;

[0227] Xaa at position 93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

[0228] Xaa at position 95 is His, Pro, Arg, Val, Leu, Gly, Asn, Ile,Phe, Ser or Thr;

[0229] Xaa at position 96 is Pro or Tyr;

[0230] Xaa at position 97 is Ile, Val, or Ala;

[0231] Xaa at position 98 is His, Ile, Asn, Leu, Asp, Ala, Thr, Leu,Arg, Gln, Glu, lys, Met, Ser, Tyr, Val or Pro;

[0232] Xaa at position 99 is Ile, Leu, Val, or Phe;

[0233] Xaa at position 100 is Lys, Leu, His, Arg, Ile, Gln, Pro, or Ser;

[0234] Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val, Asn,Ile, Leu or Tyr;

[0235] Xaa at position 102 is Gly, Glu, Lys, or Ser;

[0236] Xaa at position 104 is Trp, Val, Tyr, Met, or Leu;

[0237] Xaa at position 105 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,Leu, Lys, Ile, Asp, or His;

[0238] Xaa at position 106 is Glu, Ser, Ala, or Gly;

[0239] Xaa at position 108 is Arg, Ala, Gln, Ser or Lys;

[0240] Xaa at position 109 is Arg, Thr, Glu, Leu, Ser, or Gly;

[0241] Xaa at position 112 is Thr, Val, Gln, Glu, His, or Ser;

[0242] Xaa at position 114 is Tyr or Trp;

[0243] Xaa at position 115 is Leu or Ala;

[0244] Xaa at position 116 is Lys, Thr, Met, Val, Trp, Ser, Leu, Ala,Asn, Gln, His, Met, Phe, Tyr or Ile;

[0245] Xaa at position 117 is Thr, Ser, or Asn;

[0246] Xaa at position 119 is Glu, Ser, Pro, Leu, Thr, or Tyr;

[0247] Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;

[0248] Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;

[0249] Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys;

[0250] Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

[0251] and which can additionally have Met- preceding the amino acid inposition 1; and wherein from 1 to 14 amino acids can be deleted from theN-terminus and/or from 1 to 15 amino acids can be deleted from theC-terminus; and wherein from 4 to 44 of the amino acids designated byXaa are different from the corresponding amino acids of native (1-133)human interleukin 3.

[0252] Included in the present invention are human interleukin-3 mutantpolypeptide of the Formula III: Ala Pro Met Thr Gln Thr Thr Ser Leu LysThr Ser Trp Val Asn [SEQ ID NO:17] 1           5               10                   15 Cys Xaa Xaa Xaa IleXaa Glu Xaa Xaa Xaa Xaa Leu Lys Xaa Xaa             20              25             30 Xaa Xaa Xaa Xaa Xaa AspXaa Xaa Asn Leu Asn Xaa Glu Xaa Xaa             35              40             45 Xaa Ile Leu Met Xaa XaaAsn Leu Xaa Xaa Xaa Asn Leu Glu Xaa             50              55             60 Phe Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Asn Xaa Xaa Xaa Ile Glu             65              70             75 Xaa Xaa Leu Xaa Xaa LeuXaa Xaa Cys Xaa Pro Xaa Xaa Thr Ala             80              85             90 Xaa Pro Xaa Arg Xaa XaaXaa Xaa Xaa Xaa Xaa Gly Asp Xaa Xaa             95             100            105 Xaa Phe Xaa Xaa Lys LeuXaa Phe Xaa Xaa Xaa Xaa Leu Glu Xaa            110             115            120 Xaa Xaa Xaa Gln Gln ThrThr Leu Ser Leu Ala Ile Phe             125            130

[0253] wherein

[0254] Xaa at position 17 is Ser, Gly, Asp, Met, or Gln;

[0255] Xaa at position 18 is Asn, His, or Ile;

[0256] Xaa at position 19 is Met or Ile;

[0257] Xaa at position 21 is Asp or Glu;

[0258] Xaa at position 23 is Ile, Ala, Leu, or Gly;

[0259] Xaa at position 24 is Ile, Val, or Leu;

[0260] Xaa at position 25 is Thr, His, Gln, or Ala;

[0261] Xaa at position 26 is His or Ala;

[0262] Xaa at position 29 is Gln, Asn, or Val;

[0263] Xaa at position 30 is Pro, Gly, or Gln;

[0264] Xaa at position 31 is Pro, Asp, Gly, or Gln;

[0265] Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

[0266] Xaa at position 33 is Pro or Glu;

[0267] Xaa at position 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln,Glu, Ile, Phe, Thr or Met;

[0268] Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val;

[0269] Xaa at position 37 is Phe, Ser, Pro, or Trp;

[0270] Xaa at position 38 is Asn or Ala;

[0271] Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu,Met, Tyr or Arg;

[0272] Xaa at position 44 is Asp or Glu;

[0273] Xaa at position 45 is Gln, Val, Met, Leu, Thr, Ala, Asn, Glu, Seror Lys;

[0274] Xaa at position 46 is Asp, Phe, Ser, Thr, Ala, Asn Gln, Glu, His,Ile, Lys, Tyr, Val or Cys;

[0275] Xaa at position 50 is Glu, Ala, Asn, Ser or Asp;

[0276] Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, or His;

[0277] Xaa at position 54 is Arg or Ala;

[0278] Xaa at position 54 is Arg or Ala;

[0279] Xaa at position 55 is Arg, Thr, Val, Leu, or Gly;

[0280] Xaa at position 56 is Pro, Gly, Ser, Gln, Ala, Arg, Asn, Glu,Leu, Thr, Val or Lys;

[0281] Xaa at position 60 is Ala or Ser;

[0282] Xaa at position 62 is Asn, Pro, Thr, or Ile;

[0283] Xaa at position 63 is Arg or Lys;

[0284] Xaa at position 64 is Ala or Asn;

[0285] Xaa at position 65 is Val or Thr;

[0286] Xaa at position 66 is Lys or Arg;

[0287] Xaa at position 67 is Ser, Phe, or His;

[0288] Xaa at position 68 is Leu, Ile, Phe, or His;

[0289] Xaa at position 69 is Gln, Ala, Pro, Thr, Gln, Arg, or Gly;

[0290] Xaa at position 71 is Ala, Pro, or Arg;

[0291] Xaa at position 72 is Ser, Glu, Arg, or Asp;

[0292] Xaa at position 73 is Ala or Leu;

[0293] Xaa at position 76 is Ser, Val, Ala, Asn, Glu, Pro, or Gly;

[0294] Xaa at position 77 is Ile or Leu;

[0295] Xaa at position 79 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, orAsp;

[0296] Xaa at position 80 is Asn, Gly, Glu, or Arg;

[0297] Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu,His, Ile, Met, Phe, Ser, Thr, Tyr or Val;

[0298] Xaa at position 83 is Pro or Thr;

[0299] Xaa at position 85 is Leu or Val;

[0300] Xaa at position 87 is Leu or Ser;

[0301] Xaa at position 88 is Ala or Trp;

[0302] Xaa at position 91 is Ala or Pro;

[0303] Xaa at position 93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

[0304] Xaa at position 95 is His, Pro, Arg, Val, Leu, Gly, Asn, Phe, Seror Thr;

[0305] Xaa at position 96 is Pro or Tyr;

[0306] Xaa at position 97 is Ile or Val;

[0307] Xaa at position 98 is His, Ile, Asn, Leu, Ala, Thr, Leu, Arg,Gln, Leu, Lys, Met, Ser, Tyr, Val or Pro;

[0308] Xaa at position 99 is Ile, Leu, or Val;

[0309] Xaa at position 100 is Lys, Arg, Ile, Gln, Pro, or Ser;

[0310] Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Pro, Asn,Ile, Leu or Tyr;

[0311] Xaa at position 104 is Trp or Leu;

[0312] Xaa at position 105 is Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu,Lys, Ile, Asp, or His;

[0313] Xaa at position 106 is Glu or Gly;

[0314] Xaa at position 108 is Arg, Ala, or Ser;

[0315] Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser;

[0316] Xaa at position 112 is Thr, Val, or Gln;

[0317] Xaa at position 114 is Tyr or Trp;

[0318] Xaa at position 115 is Leu or Ala;

[0319] Xaa at position 116 is Lys, Thr, Val, Trp, Ser, Ala, His, Met,Phe, Tyr or Ile;

[0320] Xaa at position 117 is Thr or Ser;

[0321] Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;

[0322] Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Asp, or Gly;

[0323] Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys;

[0324] Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

[0325] and which can additionally have Met- preceding the amino acid inposition 1; and wherein from 1 to 14 amino acids can be deleted from theN-terminus and/or from 1 to 15 amino acids can be deleted from theC-terminus; and wherein from 4 to 35 of the amino acids designated byXaa are different from the corresponding amino acids of native(1-133)human interleukin-3.

[0326] Included in the present invention arehuman interleukin-3 mutantpolypeptide of the Formula IV: Ala Pro Met Thr Gln Thr Thr Ser Leu LysThr Ser Trp Val Asn [SEQ ID NO:18]            15              10             15 Cys Xaa Xaa Met Ile AspGlu Xaa Ile Xaa Xaa Leu Lys Xaa Xaa             20              25             30 Pro Xaa Pro Xaa Xaa AspPhe Xaa Asn Leu Asn Xaa Glu Asp Xaa             35             40              45 Xaa Ile Leu Met Xaa XaaAsn Leu Arg Xaa Xaa Asn Leu Glu Ala             50             55              60 Phe Xaa Arg Xaa Xaa LysXaa Xaa Xaa Asn Ala Ser Ala Ile Glu             65             70              75 Xaa Xaa Leu Xaa Xaa LeuXaa Pro Cys Leu Pro Xaa Xaa Thr Ala             80             85              90 Xaa Pro Xaa Arg Xaa ProIle Xaa Xaa Xaa Xaa Gly Asp Trp Xaa             95            100             105 Glu Phe Xaa Xaa Lys LeuXaa Phe Tyr Leu Xaa Xaa Leu Glu Xaa            110            115             120 Xaa Xaa Xaa Gln Gln ThrThr Leu Ser Leu Ala Ile Phe             125            130

[0327] wherein

[0328] Xaa at position 17 is Ser, Gly, Asp, or Gln;

[0329] Xaa at position 18 is Asn, His, or Ile;

[0330] Xaa at position 23 is Ile, Ala, Leu, or Gly;

[0331] Xaa at position 25 is Thr, His, or Gln;

[0332] Xaa at position 26 is His or Ala;

[0333] Xaa at position 29 is Gln or Asn;

[0334] Xaa at position 30 is Pro or Gly;

[0335] Xaa at position 32 is Leu, Arg, Asn, or Ala;

[0336] Xaa at position 34 is Leu, Val, Ser, Ala, Arg, Gln, Glu, Ile,Phe, Thr, or Met;

[0337] Xaa at position 35 is Leu, Ala, Asn, or Pro;

[0338] Xaa at position 38 is Asn or Ala;

[0339] Xaa at position 42 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyror Arg;

[0340] Xaa at position 45 is Gln, Val, Met, Leu, Ala, Asn, Glu, or Lys;

[0341] Xaa at position 46 is Asp, Phe, Ser, Gln, Glu, His, Val or Thr;

[0342] Xaa at position 50 is Glu Asn, Ser or Asp;

[0343] Xaa at position 51 is Asn, Arg, Pro, Thr, or His;

[0344] Xaa at position 55 is Arg, Leu, or Gly;

[0345] Xaa at position 56 is Pro, Gly, Ser, Ala, Asn, Val, Leu or Gln;

[0346] Xaa at position 62 is Asn, Pro, or Thr;

[0347] Xaa at position 64 is Ala or Asn;

[0348] Xaa at position 65 is Val or Thr;

[0349] Xaa at position 67 is Ser or Phe;

[0350] Xaa at position 68 is Leu or Phe;

[0351] Xaa at position 69 is Gln, Ala, Glu, or Arg;

[0352] Xaa at position 76 is Ser, Val, Asn, Pro, or Gly;

[0353] Xaa at position 77 is Ile or Leu;

[0354] Xaa at position 79 is Lys, Gly, Asn, Met, Arg, Ile, or Gly;

[0355] Xaa at position 80 is Asn, Gly, Glu, or Arg;

[0356] Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His,Met, Phe, Ser, Thr, Tyr or Val;

[0357] Xaa at position 87 is Leu or Ser;

[0358] Xaa at position 88 is Ala or Trp;

[0359] Xaa at position 91 is Ala or Pro;

[0360] Xaa at position 93 is Thr, Asp, or Ala;

[0361] Xaa at position 95 is His, Pro, Arg, Val, Gly, Asn, Ser or Thr;

[0362] Xaa at position 98 is His, Ile, Asn, Ala, Thr, Gln, Glu, Lys,Met, Ser, Tyr, Val or Leu;

[0363] Xaa at position 99 is Ile or Leu;

[0364] Xaa at position 100 is Lys or Arg;

[0365] Xaa at position 101 is Asp, Pro, Met, Lys, Thr, His, Pro, Asn,Ile, Leu or Tyr;

[0366] Xaa at position 105 is Asn, Pro, Ser, Ile or Asp;

[0367] Xaa at position 108 is Arg, Ala, or Ser;

[0368] Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser;

[0369] Xaa at position 112 is Thr or Gln;

[0370] Xaa at position 116 is Lys, Val, Trp, Ala, His, Phe, Tyr or Ile;

[0371] Xaa at position 117 is Thr or Ser;

[0372] Xaa at position 120 is Asn, Pro, Leu, His, Val, or Gln;

[0373] Xaa at position 121 is Ala, Ser, Ile, Pro, or Asp;

[0374] Xaa at position 122 is Gln, Met, Trp, Phe, Pro, His, Ile, or Tyr;

[0375] Xaa at position 123 is Ala, Met, Glu, Ser, or Leu;

[0376] and which can additionally have Met- preceding the amino acid inposition 1; and wherein from 1 to 14 amino acids can be deleted from theN-terminus and/or from 1 to 15 amino acids can be deleted from theC-terminus; and wherein from 4 to 44 of the amino acids designated byXaa are different from the corresponding amino acids of native(1-133)human interleukin-3.

[0377] Included in the present invention are (15-125)human interleukin-3mutant polypeptides of the Formula V: Asn Cys Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa [SEQ ID NO:19] 15          10              15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa AsnXaa Xaa Xaa Xaa Xaa              20              25             30 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             35              40             45 Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             50              55             60 Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             65              70             75 Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             80              85             90 Xaa Xaa Phe Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             95            100             105 Xaa Xaa Xaa Xaa Gln Gln             110

[0378] wherein

[0379] Xaa at position 3 is Ser, Lys, Gly, Asp, Met, Gln, or Arg;

[0380] Xaa at position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

[0381] Xaa at position 5 is Met, Phe, Ile, Arg, Gly, Ala, or Cys;

[0382] Xaa at position 6 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala;

[0383] Xaa at position 7 is Asp, Phe, Lys, Arg, Ala, Gly, Glu, Gln, Asn,Thr, Ser or Val;

[0384] Xaa at position 8 is Glu, Trp, Pro, Ser, Ala, His, Asp, Asn, Gln,Leu, Val, or Gly;

[0385] Xaa at position 9 is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe, Leu,Ser, or Arg;

[0386] Xaa at position 10 is Ile, Gly, Val, Arg, Ser, Phe, or Leu;

[0387] Xaa at position 11 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

[0388] Xaa at position 12 is His, Thr, Phe, Gly, Arg, Ala, or Trp;

[0389] Xaa at position 13 is Leu, Gly, Arg, Thr, Ser, or Ala;

[0390] Xaa at position 14 is Lys, Arg, Leu, Gln, Gly, Pro, Val or Trp;

[0391] Xaa at position 15 is Gln, Asn, Leu, Pro, Arg, or Val;

[0392] Xaa at position 16 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, orLys;

[0393] Xaa at position 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

[0394] Xaa at position 18 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu;

[0395] Xaa at position 19 is Pro, Leu, Gln, Ala, Thr, or Glu;

[0396] Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Glu, Gln, Thr,Arg, Ala, Phe, Ile or Met;

[0397] Xaa at position 21 is Leu, Ala, Gly, Asn, Pro, Gln, or Val;

[0398] Xaa at position 22 is Asp, Leu, or Val;

[0399] Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile;

[0400] Xaa at position 24 is Asn, or Ala;

[0401] Xaa at position 26 is Leu, Trp, or Arg;

[0402] Xaa at position 27 is Asn, Cys, Arg, Leu, His, Met, Pro;

[0403] Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Lys, Asn, Thr,Leu, Val, Glu, Phe, Tyr, Ile or Met;

[0404] Xaa at position 29 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys,Gln, Arg, Thr, Gly or Ser;

[0405] Xaa at position 30 is Asp, Ser, Leu, Arg, Lys, Thr, Met, Trp,Glu, Asn, Gln, Ala or Pro;

[0406] Xaa at position 31 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys,Asp, Asn, Arg, Ser, Ala, Ile, Glu, His or Trp;

[0407] Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Glu, Asn, Gln,Lys, His, Ala, Tyr, Ile, Val or Gly;

[0408] Xaa at position 33 is Ile, Gly, Val, Ser, Arg, Pro, or His;

[0409] Xaa at position 34 is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu,Lys, Thr, Ala, Met, Val or Asn;

[0410] Xaa at position 35 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;

[0411] Xaa at position 36 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser,Ala, Ile, Val, His, Phe, Met or Gln;

[0412] Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;

[0413] Xaa at position 38 is Asn, His, Arg, Leu, Gly, Ser, or Thr;

[0414] Xaa at position 39 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser,Met, or;

[0415] Xaa at position 40 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn,Lys, His, Ala or Leu;

[0416] Xaa at position 41 is Arg, Thr, Val, Ser, Leu, or Gly;

[0417] Xaa at position 42 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His,Thr, Ala, Tyr, Phe, Leu, Val or Lys;

[0418] Xaa at position 43 is Asn or Gly;

[0419] Xaa at position 44 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;

[0420] Xaa at position 45 is Glu Tyr, His, Leu, Pro, or Arg;

[0421] Xaa at position 46 is Ala, Ser, Pro, Tyr, Asn, or Thr;

[0422] Xaa at position 47 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser;

[0423] Xaa at position 48 is Asn, His, Val, Arg, Pro, Thr, Asp, or Ile;

[0424] Xaa at position 49 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or Val;

[0425] Xaa at position 50 is Ala, Asn, Pro, Ser, or Lys;

[0426] Xaa at position 51 is Val, Thr, Pro, His, Leu, Phe, or Ser;

[0427] Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

[0428] Xaa at position 53 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, orHis;

[0429] Xaa at position 54 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or His;

[0430] Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly, orLeu;

[0431] Xaa at position 56 is Asn, Leu, Val, Trp, Pro, or Ala;

[0432] Xaa at position 57 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln,Trp, or Asn;

[0433] Xaa at position 58 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;

[0434] Xaa at position 59 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or Arg;

[0435] Xaa at position 60 is Ile, Met, Thr, Pro, Arg, Gly, Ala;

[0436] Xaa at position 61 is Glu, Lys, Gly, Asp, Pro, Trp, Arg, Ser,Gln, or Leu;

[0437] Xaa at position 62 is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly, orAsp;

[0438] Xaa at position 63 is Ile, Ser, Arg, Thr, or Leu;

[0439] Xaa at position 64 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg;

[0440] Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or Asp;

[0441] Xaa at position 66 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or Arg;

[0442] Xaa at position 67 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys;

[0443] Xaa at position 68 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn,His, Thr, Ser, Ala, Tyr, Phe, Ile, Met or Val;

[0444] Xaa at position 69 is Pro, Ala, Thr, Trp, Arg, or Met;

[0445] Xaa at position 70 is Cys, Glu, Gly, Arg, Met, or Val;

[0446] Xaa at position 71 is Leu, Asn, Val, or Gln;

[0447] Xaa at position 72 is Pro, Cys, Arg, Ala, or Lys;

[0448] Xaa at position 73 is Leu, Ser, Trp, or Gly;

[0449] Xaa at position 74 is Ala, Lys, Arg, Val, or Trp;

[0450] Xaa at position 75 is Thr, Asp, Cys, Leu, Val, Glu, His, Asn, orSer;

[0451] Xaa at position 76 is Ala, Pro, Ser, Thr, Gly, Asp, Ile, or Met;

[0452] Xaa at position 77 is Ala, Pro, Ser, Thr, Phe, Leu, Asp, or His;

[0453] Xaa at position 78 is Pro, Phe, Arg, Ser, Lys, His, Ala, Gly, Ileor Leu;

[0454] Xaa at position 79 is Thr, Asp, Ser, Asn, Pro, Ala, Leu, or Arg;

[0455] Xaa at position 80 is Arg, Ile, Ser, Glu, Leu, Val, Gln, Lys,His, Ala or Pro;

[0456] Xaa at position 81 is His, Gln, Pro, Arg, Val, Leu, Gly, Thr,Asn, Lys, Ser, Ala, Trp, Phe, Ile or Tyr;

[0457] Xaa at position 82 is Pro, Lys, Tyr, Gly, Ile, or Thr;

[0458] Xaa at position 83 is Ile, Val, Lys, Ala, or Asn;

[0459] Xaa at position 84 is His, Ile, Asn, Leu, Asp, Ala, Thr, Glu,Gln, Ser, Phe, Met, Val, Lys, Arg, Tyr or Pro;

[0460] Xaa at position 85 is Ile, Leu, Arg, Asp, Val, Pro, Gln, Gly,Ser, Phe, or His;

[0461] Xaa at position 86 is Lys, Tyr, Leu, His, Arg, Ile, Ser, Gln,Pro;

[0462] Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Val, Tyr,Glu, Asn, Ser, Ala, Gly, Ile, Leu or Gln;

[0463] Xaa at position 88 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro;

[0464] Xaa at position 89 is Asp, or Ser;

[0465] Xaa at position 90 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu,Gln, Lys, Ala, Phe, or Gly;

[0466] Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,Leu, Lys, Ile, Asp, or His;

[0467] Xaa at position 92 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro;

[0468] Xaa at position 94 is Arg, Lys, Asp, Leu, Thr, Ile, Gln, His,Ser, Ala, or Pro;

[0469] Xaa at position 95 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly;

[0470] Xaa at position 96 is Lys, Asn, Thr, Leu, Gln, Arg, His, Glu,Ser, Ala or Trp;

[0471] Xaa at position 97 is Leu, Ile, Arg, Asp, or Met;

[0472] Xaa at position 98 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe;

[0473] Xaa at position 99 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp,Lys, Leu, Ile, Val or Asn;

[0474] Xaa at position 100 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;

[0475] Xaa at position 101 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr,Trp, or Met;

[0476] Xaa at position 102 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu,Arg, Trp, Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile;

[0477] Xaa at position 103 is Thr, Ser, Asn, Ile, Trp, Lys, or Pro;

[0478] Xaa at position 104 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;

[0479] Xaa at position 105 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg;

[0480] Xaa at position 106 is Asn, Ala, Pro, Leu, His, Val, or Gln;

[0481] Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;

[0482] Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys;

[0483] Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

[0484] and which can additionally have Met- or Met-Ala- preceding theamino acid in position 1; and wherein from 4 to 44 of the amino acidsdesignated by Xaa are different from the corresponding native aminoacids of (1-133) human interleukin-3; or a polypeptide havingsubstantially the same structure and substantially the same biologicalactivity.

[0485] Included in the present invention are (15-125)human interleukin-3mutant polypeptides of the Formula VI: Asn Cys Xaa Xaa Xaa Xaa Xaa GluXaa Xaa Xaa Xaa Leu Xaa Xaa [SEQ ID NO:20] 1               5                   10                  15 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa                 20                  25                  30 Xaa Xaa XaaLeu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa                 35                  40                  45 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa                 50                  55                  60 Xaa Xaa XaaLeu Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa                 65                  70                  75 Xaa Xaa XaaXaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa                 80                  85                  90 Xaa Xaa PheXaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa                 95                 100                 105 Xaa Xaa XaaXaa Gln Gln                 110

[0486] wherein

[0487] Xaa at position 3 is Ser, Gly, Asp, Met, or Gln;

[0488] Xaa at position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln;

[0489] Xaa at position 5 is Met, Phe, Ile, Arg, or Ala;

[0490] Xaa at position 6 is Ile or Pro;

[0491] Xaa at position 7 is Asp, or Glu;

[0492] Xaa at position 9 is Ile, Val, Ala, Leu, or Gly;

[0493] Xaa at position 10 is Ile, Val, Phe, or Leu;

[0494] Xaa at position 11 is Thr, His, Gly, Gln, Arg, Pro, or Ala;

[0495] Xaa at position 12 is His, Phe, Gly, Arg, or Ala;

[0496] Xaa at position 14 is Lys, Leu, Gln, Gly, Pro, or Val;

[0497] Xaa at position 15 is Gln, Asn, Leu, Arg, or Val;

[0498] Xaa at position 16 is Pro, His, Thr, Gly, or Gln;

[0499] Xaa at position 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln;

[0500] Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

[0501] Xaa at position 19 is Pro, Leu, Gln, Ala, or Glu;

[0502] Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln,Glu, Ile, Phe, Thr or Met;

[0503] Xaa at position 21 is Leu, Ala, Asn, Pro, Gln, or Val;

[0504] Xaa at position 22 is Asp or Leu;

[0505] Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile;

[0506] Xaa at position 24 is Asn or Ala;

[0507] Xaa at position 27 is Asn, Cys, Arg, His, Met, or Pro;

[0508] Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu,Met, Tyr, or Arg;

[0509] Xaa at position 30 is Asp, or Glu;

[0510] Xaa at position 31 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn Glu,Ser or Trp;

[0511] Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln,Glu, His, Ile, Lys, Tyr, Val or Gly;

[0512] Xaa at position 33 is Ile, Val, or His;

[0513] Xaa at position 35 is Met, Asn, or Asp;

[0514] Xaa at position 36 is Glu, Thr, Ala, Asn, Ser or Asp;

[0515] Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;

[0516] Xaa at position 38 is Asn or Gly;

[0517] Xaa at position 39 is Leu, Met, or Phe;

[0518] Xaa at position 40 is Arg, Ala or Ser;

[0519] Xaa at position 41 is Arg, Thr, Val, Leu, or Gly;

[0520] Xaa at position 42 is Pro, Gly, Cys, Ser, Gln, Ala, Arg, Asn,Glu, His, Leu, Thr, Val or Lys;

[0521] Xaa at position 45 is Glu, Tyr, His, Leu, or Arg;

[0522] Xaa at position 46 is Ala, Ser, Asn, or Thr;

[0523] Xaa at position 47 is Phe or Ser;

[0524] Xaa at position 48 is Asn, Val, Pro, Thr, or Ile;

[0525] Xaa at position 49 is Arg, Tyr, Lys, Ser, His, or Val;

[0526] Xaa at position 50 is Ala or Asn;

[0527] Xaa at position 51 is Val, Thr, Leu, or Ser;

[0528] Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;

[0529] Xaa at position 53 is Ser, Phe, Val, Gly, Asn, Ile, or His;

[0530] Xaa at position 54 is Leu, Val, Ile, Phe, or His;

[0531] Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

[0532] Xaa at position 56 is Asn or Pro;

[0533] Xaa at position 57 is Ala, Met, Pro, Arg, Glu, Thr, or Gln;

[0534] Xaa at position 58 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;

[0535] Xaa at position 59 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, Arg, orPro;

[0536] Xaa at position 60 is Ile or Met;

[0537] Xaa at position 61 is Glu, Gly, Asp, Ser, or Gln;

[0538] Xaa at position 62 is Ser, Val, Ala, Asn, Glu, Pro, Gly, or Asp;

[0539] Xaa at position 63 is Ile, Ser, or Leu;

[0540] Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or Asp;

[0541] Xaa at position 66 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;

[0542] Xaa at position 67 is Leu, or Val;

[0543] Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu,His, Met, Phe, Ser, Thr, Tyr or Val;

[0544] Xaa at position 69 is Pro, Ala, Thr, Trp, or Met;

[0545] Xaa at position 71 is Leu or Val;

[0546] Xaa at position 73 is Leu or Ser;

[0547] Xaa at position 74 is Ala, Arg, or Trp;

[0548] Xaa at position 75 is Thr, Asp, Glu, His, Asn, or Ser;

[0549] Xaa at position 76 is Ala, Asp, or Met;

[0550] Xaa at position 77 is Ala, Pro, Ser, Thr, Phe, Leu, or Asp;

[0551] Xaa at position 78 is Pro or Ser;

[0552] Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

[0553] Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Ile,Phe, Ser or Thr;

[0554] Xaa at position 82 is Pro or Tyr;

[0555] Xaa at position 83 is Ile, Val, or Ala;

[0556] Xaa at position 84 is His, Ile, Asn, Leu, Asp, Ala, Thr, Arg,Gln, Glu, Lys, Met, Ser, Tyr, Val or Pro;

[0557] Xaa at position 85 is Ile, Leu, Val, or Phe;

[0558] Xaa at position 86 is Lys, Leu, His, Arg, Ile, Gln, Pro or Ser;

[0559] Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Val, Asn,Ile, Leu or Tyr;

[0560] Xaa at position 88 is Gly, Glu, Lys, or Ser;

[0561] Xaa at position 90 is Trp, Val, Tyr, Met, or Leu;

[0562] Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,Leu, Lys, Ile, Asp, or His;

[0563] Xaa at position 92 is Glu, Ser, Ala, or Gly;

[0564] Xaa at position 94 is Arg, Ala, Gln, Ser or Lys;

[0565] Xaa at position 95 is Arg, Thr, Glu, Leu, Ser, or Gly;

[0566] Xaa at position 98 is Thr, Val, Gln, Glu, His, or Ser;

[0567] Xaa at position 100 is Tyr or Trp;

[0568] Xaa at position 101 is Leu or Ala;

[0569] Xaa at position 102 is Lys, Thr, Met, Val, Trp, Ser, Leu, Ala,Asn, Gln, His, Met, Phe, Tyr or Ile;

[0570] Xaa at position 103 is Thr, Ser, or Asn;

[0571] Xaa at position 105 is Glu, Ser, Pro, Leu, Thr, or Tyr;

[0572] Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;

[0573] Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly;

[0574] Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys;

[0575] Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

[0576] and which can additionally have Met- or Met-Ala- preceding theamino acid in position 1; and wherein from 4 to 44 of the amino acidsdesignated by Xaa are different from the corresponding amino acids ofnative (1-133) human interleukin-3; or a polypeptide havingsubstantially the same structure and substantially the same biologicalactivity.

[0577] Included in the present invention are (15-125)human interleukin-3mutant polypeptides of the Formula VII: Asn Cys Xaa Xaa Xaa Ile Xaa GluXaa Xaa Xaa Xaa Leu Lys Xaa [SEQ ID NO:21] 1               5                   10                 15 Xaa Xaa XaaXaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa                 20                  25                  30 Xaa Xaa IleLeu Met Xaa Xaa Asn Leu Xaa Xaa Xaa Asn Leu Glu                 35                  40                  45 Xaa Phe XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile                 50                  55                  60 Glu Xaa XaaLeu Xaa Xaa Leu Xaa Xaa Cys Xaa Pro Xaa Xaa Thr                 65                  70                  75 Ala Xaa ProXaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa                 80                  85                  90 Xaa Xaa PheXaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Glu                 95                  100                105 Xaa Xaa XaaXaa Gln Gln                 110

[0578] wherein

[0579] Xaa at position 3 is Ser, Gly, Asp, Met, or Gln;

[0580] Xaa at position 4 is Asn, His, or Ile;

[0581] Xaa at position 5 is Met or Ile;

[0582] Xaa at position 7 is Asp or Glu;

[0583] Xaa at position 9 is Ile, Ala, Leu, or Gly;

[0584] Xaa at position 10 is Ile, Val, or Leu;

[0585] Xaa at position 11 is Thr, His, Gln, or Ala;

[0586] Xaa at position 12 is His or Ala;

[0587] Xaa at position 15 is Gln, Asn, or Val;

[0588] Xaa at position 16 is Pro, Gly, or Gln;

[0589] Xaa at position 17 is Pro, Asp, Gly, or Gln;

[0590] Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;

[0591] Xaa at position 19 is Pro or Glu;

[0592] Xaa at position 20 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln,Glu, Ile, Phe, Thr or Met;

[0593] Xaa at position 21 is Leu, Ala, Asn, Pro, Gln, or Val;

[0594] Xaa at position 23 is Phe, Ser, Pro, or Trp;

[0595] Xaa at position 24 is Asn or Ala;

[0596] Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, MetTyr or Arg;

[0597] Xaa at position 30 is Asp or Glu;

[0598] Xaa at position 31 is Gln, Val, Met, Leu, Thr, Ala, Asn, Glu, Seror Lys;

[0599] Xaa at position 32 is Asp, Phe, Ser, Thr, Ala, Asn, Gln, Glu,His, Ile, Lys, Tyr, Val or Cys;

[0600] Xaa at position 36 is Glu, Ala, Asn, Ser or Asp;

[0601] Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His;

[0602] Xaa at position 40 is Arg or Ala;

[0603] Xaa at position 41 is Arg, Thr, Val, Leu, or Gly;

[0604] Xaa at position 42 is Pro, Gly, Ser, Gln, Ala, Arg, Asn, Glu,Leu, Thr, Val or Lys;

[0605] Xaa at position 46 is Ala or Ser;

[0606] Xaa at position 48 is Asn, Pro, Thr, or Ile;

[0607] Xaa at position 49 is Arg or Lys;

[0608] Xaa at position 50 is Ala or Asn;

[0609] Xaa at position 51 is Val or Thr;

[0610] Xaa at position 52 is Lys or Arg;

[0611] Xaa at position 53 is Ser, Phe, or His;

[0612] Xaa at position 54 is Leu, Ile, Phe, or His;

[0613] Xaa at position 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly;

[0614] Xaa at position 57 is Ala, Pro, or Arg;

[0615] Xaa at position 58 is Ser, Gln, Arg, or Asp;

[0616] Xaa at position 59 is Ala or Leu;

[0617] Xaa at position 62 is Ser, Val, Ala, Asn, Glu, Pro, or Gly;

[0618] Xaa at position 63 is Ile or Leu;

[0619] Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, orAsp;

[0620] Xaa at position 66 is Asn, Gly, Glu, or Arg;

[0621] Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Ala, Asn, Glu,His, Ile, Met, Phe, Ser, Thr, Tyr or Val;

[0622] Xaa at position 69 is Pro or Thr;

[0623] Xaa at position 71 is Leu or Val;

[0624] Xaa at position 73 is Leu or Ser;

[0625] Xaa at position 74 is Ala or Trp;

[0626] Xaa at position 77 is Ala or Pro;

[0627] Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;

[0628] Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Phe, Seror Thr;

[0629] Xaa at position 82 is Pro or Tyr;

[0630] Xaa at position 83 is Ile or Val;

[0631] Xaa at position 84 is His, Ile, Asn, Leu, Ala, Thr, Leu, Arg,Gln, Leu, Lys, Met, Ser, Tyr, Val or Pro;

[0632] Xaa at position 85 is Ile, Leu, or Val;

[0633] Xaa at position 86 is Lys, Arg, Ile, Gln, Pro, or Ser;

[0634] Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr, Asn, Ile, Leuor Tyr;

[0635] Xaa at position 90 is Trp or Leu;

[0636] Xaa at position 91 is Asn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu,Lys, Ile, Asp, or His;

[0637] Xaa at position 92 is Glu, or Gly;

[0638] Xaa at position 94 is Arg, Ala, or Ser;

[0639] Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser;

[0640] Xaa at position 98 is Thr, Val, or Gln;

[0641] Xaa at position 100 is Tyr or Trp;

[0642] Xaa at position 101 is Leu or Ala;

[0643] Xaa at position 102 is Lys, Thr, Val, Trp, Ser, Ala, His, Met,Phe, Tyr or Ile;

[0644] Xaa at position 103 is Thr or Ser;

[0645] Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;

[0646] Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Asp, or Gly;

[0647] Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys;

[0648] Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu;

[0649] which can additionally have Met- or Met-Ala- preceding the aminoacid in position 1; and wherein from 4 to 35 of the amino acidsdesignated by Xaa are different from the corresponding amino acids ofnative human interleukin-3.

[0650] Included in the present invention are (15-125)human interleukin-3mutant polypeptides of the Formula VIII: Asn Cys Xaa Xaa Met Ile Asp GluXaa Ile Xaa Xaa Leu Lys Xaa [SEQ ID NO:22] 1               5                   10                  15 Xaa Pro XaaPro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Glu Asp                 20                  25                  30 Xaa Xaa IleLeu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Glu                 35                  40                  45 Ala Phe XaaArg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile                 50                  55                  60 Glu Xaa XaaLeu Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr                 65                  70                  75 Ala Xaa ProXaa Arg Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp                 80                  85                  90 Xaa Glu PheXaa Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu                 95                 100                 105 Xaa Xaa XaaXaa Gln Gln                 110

[0651] wherein

[0652] Xaa at position 3 is Ser, Gly, Asp, or Gln;

[0653] Xaa at position 4 is Asn, His, or Ile;

[0654] Xaa at position 9 is Ile, Ala, Leu, or Gly;

[0655] Xaa at position 11 is Thr, His, or Gln;

[0656] Xaa at position 12 is His or Ala;

[0657] Xaa at position 15 is Gln or Asn;

[0658] Xaa at position 16 is Pro or Gly;

[0659] Xaa at position 18 is Leu, Arg, Asn, or Ala;

[0660] Xaa at position 20 is Leu, Val, Ser, Ala, Arg, Gln, Glu, Ile,Phe, Thr or Met;

[0661] Xaa at position 21 is Leu, Ala, Asn, or Pro;

[0662] Xaa at position 24 is Asn or Ala;

[0663] Xaa at position 28 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyror Arg;

[0664] Xaa at position 31 is Gln, Val, Met, Leu, Ala, Asn, Glu or Lys;

[0665] Xaa at position 32 is Asp, Phe, Ser, Ala, Gln, Glu, His, Val orThr;

[0666] Xaa at position 36 is Glu, Asn, Ser or Asp;

[0667] Xaa at position 37 is Asn, Arg, Pro, Thr, or His;

[0668] Xaa at position 41 is Arg, Leu, or Gly;

[0669] Xaa at position 42 is Pro, Gly, Ser, Ala, Asn, Val, Leu or Gln;

[0670] Xaa at position 48 is Asn, Pro, or Thr;

[0671] Xaa at position 50 is Ala or Asn;

[0672] Xaa at position 51 is Val or Thr;

[0673] Xaa at position 53 is Ser or Phe;

[0674] Xaa at position 54 is Leu or Phe;

[0675] Xaa at position 55 is Gln, Ala, Glu, or Arg;

[0676] Xaa at position 62 is Ser, Val, Asn, Pro, or Gly;

[0677] Xaa at position 63 is Ile or Leu;

[0678] Xaa at position 65 is Lys, Asn, Met, Arg, Ile, or Gly;

[0679] Xaa at position 66 is Asn, Gly, Glu, or Arg;

[0680] Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His,Met, Phe, Ser, Thr, Tyr or Val;

[0681] Xaa at position 73 is Leu or Ser;

[0682] Xaa at position 74 is Ala or Trp;

[0683] Xaa at position 77 is Ala or Pro;

[0684] Xaa at position 79 is Thr, Asp, or Ala;

[0685] Xaa at position 81 is His, Pro, Arg, Val, Gly, Asn, Ser or Thr;

[0686] Xaa at position 84 is His, Ile, Asn, Ala, Thr, Arg, Gln, Glu,Lys, Met, Ser, Tyr, Val or Leu;

[0687] Xaa at position 85 is Ile or Leu;

[0688] Xaa at position 86 is Lys or Arg;

[0689] Xaa at position 87 is Asp, Pro, Met, Lys, His, Pro, Asn, Ile, Leuor Tyr;

[0690] Xaa at position 91 is Asn, Pro, Ser, Ile or Asp;

[0691] Xaa at position 94 is Arg, Ala, or Ser;

[0692] Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser;

[0693] Xaa at position 98 is Thr or Gln;

[0694] Xaa at position 102 is Lys, Val, Trp, or Ile;

[0695] Xaa at position 103 is Thr, Ala, His, Phe, Tyr or Ser;

[0696] Xaa at position 106 is Asn, Pro, Leu, His, Val, or Gln;

[0697] Xaa at position 107 is Ala, Ser, Ile, Pro, or Asp;

[0698] Xaa at position 108 is Gln, Met, Trp, Phe, Pro, His, Ile, or Tyr;

[0699] Xaa at position 109 is Ala, Met, Glu, Ser, or Leu;

[0700] and which can additionally have Met- or Met-Ala- preceding theamino acid in position 1; and wherein from 4 to 26 of the amino acidsdesignated by Xaa are different from the corresponding amino acids ofnative (1-133)human interleukin-3; or a polypeptide having substantiallythe same structure and substantially the same biological activity.

[0701] The present invention includes polypeptides of the formula [SEQID NO:129]        1               5                 10 (Met)_(m)-Ala ProMet Thr Gln Thr Thr Ser Leu Lys                 15                 20Thr Ser Trp Val Asn Cys Ser Xaa Xaa Xaa Asp Glu        25                  30 Ile Ile Xaa His Leu Lys Xaa Pro Pro XaaPro Xaa 35                    40                  45  Leu Asp Xaa XaaAsn Leu Asn Xaa Glu Asp Xaa Asp        50                       55 IleLeu Xaa Glu Xaa Asn Leu Arg Xaa Xaa Asn Leu   60                   65                  70 Xaa Xaa Phe Xaa XaaAlaXaa Lys Xaa Leu Xaa Asn Ala         75                      80 Ser XaaIle Glu Xaa Ile Leu Xaa Asn Leu Xaa Pro    85                  90                  95 Cys Xaa Pro Xaa Xaa ThrAla Xaa Pro Xaa Arg Xaa                 100                  105 Pro IleXaa Ile Xaa Xaa Gly Asp Trp Xaa Glu Phe        110                  115Arg Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Glu120                125    130 Xaa Ala Gln Xaa Gln Gln Thr Thr Leu SerLeu Ala Ile Phe

[0702] wherein m is 0 or 1; Xaa at position 18 is Asn or Ile; Xaa atposition 19 is Met, Ala or Ile; Xaa at position 20 is Ile, Pro or Ile;Xaa at position 23 is Ile, Ala or Leu; Xaa at position 25 is Thr or His;Xaa at position 29 is Gln, Arg, Val or Ile; Xaa at position 32 is Leu,Ala, Asn or Arg; Xaa at position 34 is Leu or Ser; Xaa at position 37 isPhe, Pro, or Ser; Xaa at position 38 is Asn or Ala; Xaa at position 42is Gly, Ala, Ser, Asp or Asn; Xaa at position 45 is Gln, Val, or Met;Xaa at position 46 is Asp or Ser; Xaa at position 49 is Met, Ile, Leu orAsp; Xaa at position 50 is Glu or Asp; Xaa at position 51 is Asn Arg orSer; Xaa at position 55 is Arg, Leu, or Thr; Xaa at position 56 is Proor Ser; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala orSer; Xaa at position 62 is Asn, Val or Pro; Xaa at position 63 is Arg orHis; Xaa at position 65 is Val or Ser; Xaa at position 67 is Ser, Asn,His or Gln; Xaa at position 69 is Gln or Glu; Xaa at position 73 is Alaor Gly; Xaa at position 76 is Ser, Ala or Pro; Xaa at position 79 isLys, Arg or Ser; Xaa at position 82 is Leu, Glu, Val or Trp; Xaa atposition 85 is Leu or Val; Xaa at position 87 is Leu, Ser, Tyr; Xaa atposition 88 is Ala or Trp; Xaa at position 91 is Ala or Pro; Xaa atposition 93 is Pro or Ser; Xaa at position 95 is His or Thr; Xaa atposition 98 is His, Ile, or Thr; Xaa at position 100 is Lys or Arg; Xaaat position 101 is Asp, Ala or Met; Xaa at position 105 is Asn or Glu;Xaa at position 109 is Arg, Glu or Leu; Xaa at position 112 is Thr orGln; Xaa at position 116 is Lys, Val, Trp or Ser; Xaa at position 117 isThr or Ser; Xaa at position 120 is Asn, Gln, or His; Xaa at position 123is Ala or Glu; with the proviso that from four to twenty-six of theamino acids designated by Xaa are different from the corresponding aminoacids of native human interleukin-3; or a polypeptide havingsubstantially the same structure and substantially the same biologicalactivity.

[0703] Preferred polypeptides of the present invention are those of theformula [SEQ ID NO:130]              1              .5(Met_(m)-Ala_(n))p-Asn Cys Ser Xaa Xaa Xaa Asp Glu Xaa 10                  15                   20 Ile Xaa His Leu Lys Xaa ProPro Xaa Pro Xaa Leu              25                 30 Asp Xaa Xaa AsnLeu Asn Xaa Glu Asp Xaa Xaa Ile   35                   40                  45 Leu Xaa Glu Xaa Asn LeuArg Xaa Xaa Asn Leu Xaa                 50                  55 Xaa PheXaa Xaa Ala Xaa Lys Xaa Leu Xaa Asn Ala         60                  65Ser Xaa Ile Glu Xaa Ile Leu Xaa Asn Xaa Xaa Pro70                  75                  80 Cys Xaa Pro Xaa Ala Thr AlaXaa Pro Xaa Arg Xaa             85                  90 Pro Ile Xaa IleXaa Xaa Gly Asp Trp Xaa Glu Phe    95                  100                 105 Arg Xaa Lys Leu Xaa PheTyr Leu Xaa Xaa Leu Glu                  110 Xaa Ala Gln Xaa Gln Gln

[0704] wherein m is 0 or 1; n is 0 or 1; p is 0 or 1; Xaa at position 4is Asn or Ile; Xaa at position 5 is Met, Ala or Ile: Xaa at position 6is Ile, Pro or Leu; Xaa at position 9 is Ile, Ala or Leu; Xaa atposition 11 is Thr or His; Xaa at position 15 is Gln, Arg, Val or Ile;Xaa at position 18 is Leu, Ala, Asn or Arg; Xaa at position 20 is Leu orSer; Xaa at position 23 is Phe, Pro, or Ser; Xaa at position 24 is Asnor Ala; Xaa at position 28 is Gly, Ala, Ser, Asp or Asn; Xaa at position31 is Gln, Val, or Met; Xaa at position 32 is Asp or Ser; Xaa atposition 35 is Met, Ile or Asp; Xaa at position 36 is Glu or Asp; Xaa atposition 37 is Asn, Arg or Ser; Xaa at position 41 is Arg, Leu, or Thr;Xaa at position 42 is Pro or Ser; Xaa at position 45 is Glu or Leu; Xaaat position 46 is Ala or Ser; Xaa at position 48 is Asn, Val or Pro; Xaaat position 49 is Arg or His; Xaa at position 51 is Val or Ser; Xaa atposition 53 is Ser, Asn, His or Gln; Xaa at position 55 is Gln or Glu;Xaa at position 59 is Ala or Gly; Xaa at position 62 is Ser, Ala or Pro;Xaa at position 65 is Lys, Arg or Ser; Xaa at position 67 is Leu, Glu,or Val; Xaa at position 68 is Leu, Glu, Val or Trp; Xaa at position 71is Leu or Val; Xaa at position 73 is Leu, Ser or Tyr; Xaa at position 74is Ala or Trp; Xaa at position 77 is Ala or Pro; Xaa at position 79 isPro or Ser; Xaa at position 81 is His or Thr; Xaa at position 84 is His,Ile, or Thr; Xaa at position 86 is Lys or Arg; Xaa at position 87 isAsp, Ala or Met; Xaa at position 91 is Asn or Glu; Xaa at position 95 isArg, Glu, Leu; Xaa at position 98 Thr or Gln; Xaa at position 102 isLys, Val, Trp or Ser; Xaa at position 103 is Thr or Ser; Xaa at position106 is Asn, Gln, or His; Xaa at position 109 is Ala or Glu; with theproviso that from four to twenty-six of the amino acids designated byXaa are different from the corresponding amino acids of native(15-125)human interleukin-3; or a polypeptide having substantially thesame structure and substantially the same biological activity.

[0705] “Mutant amino acid sequence,” “mutant protein” or “mutantpolypeptide” refers to a polypeptide having an amino acid sequence whichvaries from a native sequence or is encoded by a nucleotide sequenceintentionally made variant from a native sequence. “Mutant protein,”“variant protein” or “mutein” means a protein comprising a mutant aminoacid sequence and includes polypeptides which differ from the amino acidsequence of native hIL-3 due to amino acid deletions, substitutions, orboth. “Native sequence” refers to an amino acid or nucleic acid sequencewhich is identical to a wild-type or native form of a gene or protein.

[0706] Human IL-3 can be characterized by its ability to stimulatecolony formation by human hematopoietic progenitor cells. The coloniesformed include erythroid, granulocyte, megakaryocyte, granulocyticmacrophages and mixtures thereof. Human IL-3 has demonstrated an abilityto restore bone marrow function and peripheral blood cell populations totherapeutically beneficial levels in studies performed initially inprimates and subsequently in humans (Gillio, A. P., et al. (1990);Ganser, A, et al. (1990); Falk, S., et al. (1991). Additional activitiesof hIL-3 include the ability to stimulate leukocyte migration andchemotaxis; the ability to prime human leukocytes to produce high levelsof inflammatory mediators like leukotrienes and histamine; the abilityto induce cell surface expression of molecules needed for leukocyteadhesion; and the ability to trigger dermal inflammatory responses andfever. Many or all of these biological activities of hIL-3 involvesignal transduction and high affinity receptor binding. Mutantpolypeptides of the present invention may exhibit useful properties suchas having similar or greater biological activity when compared to nativehIL-3 or by having improved half-life or decreased adverse side effects,or a combination of these properties. They may also be useful asantagonists. hIL-3 mutant polypeptides which have little or no activitywhen compared to native hIL-3 may still be useful as antagonists, asantigens for the production of antibodies for use in immunology orimmunotherapy, as genetic probes or as intermediates used to constructother useful hIL-3 muteins. Since hIL-3 functions by binding to itsreceptor(s) and triggering second messages resulting in competent signaltransduction, hIL-3 muteins of this invention may be useful in helpingto determine which specific amino acid sequences are responsible forthese activities.

[0707] The novel hIL-3 mutant polypeptides of the present invention willpreferably have at least one biological property of human IL-3 or of anIL-3-like growth factor and may have more than one IL-3-like biologicalproperty, or an improved property, or a reduction in an undesirablebiological property of human IL-3. Some mutant polypeptides of thepresent invention may also exhibit an improved side effect profile. Forexample, they may exhibit a decrease in leukotriene release or histaminerelease when compared to native hIL-3 or (15-125) hIL-3. Such hIL-3 orhIL-3-like biological properties may include one or more of thefollowing biological characteristics and in vivo and in vitroactivities.

[0708] One such property is the support of the growth anddifferentiation of progenitor cells committed to erythroid, lymphoid,and myeloid lineages. For example, in a standard human bone marrowassay, an IL-3-like biological property is the stimulation ofgranulocytic type colonies, megakaryocytic type colonies,monocyte/macrophage type colonies, and erythroid bursts. Other IL-3-likeproperties are the interaction with early multipotential stem cells, thesustaining of the growth of pluripotent precursor cells, the ability tostimulate chronic myelogenous leukemia (CML) cell proliferation, thestimulation of proliferation of mast cells, the ability to support thegrowth of various factor-dependent cell lines, and the ability totrigger immature bone marrow cell progenitors. Other biologicalproperties of IL-3 have been disclosed in the art. Human IL-3 also hassome biological activities which may in some cases be undesirable, forexample the ability to stimulate leukotriene release and the ability tostimulate increased histamine synthesis in spleen and bone marrowcultures and in vivo.

[0709] Biological activity of hIL-3 and hIL-3 mutant proteins of thepresent invention is determined by DNA synthesis by human acutemyelogenous leukemia cells (AML). The factor-dependent cell line AML 193was adapted for use in testing biological activity.

[0710] One object of the present invention is to provide hIL-3 muteinsand hIL-3 deletion muteins with four or more amino acid substitutions inthe polypeptide sequence which have similar or improved biologicalactivity in relation to native hIL-3 or native (15-125)hIL-3.

[0711] The present invention includes mutant polypeptides comprisingminimally amino acids residues 15 to 118 of hIL-3 with or withoutadditional amino acid extensions to the N-terminus and/or C-terminuswhich further contain four or more amino acid substitutions in the aminoacid sequence of the polypeptide. It has been found that the(15-125)hIL-3 mutant is more soluble than is hIL-3 when expressed in thecytoplasm of E. coli, and the protein is secreted to the periplasm in E.coli at higher levels compared to native hIL-3.

[0712] When expressed in the E. coli cytoplasm, the above-mentionedmutant hIL-3 polypeptides of the present invention may also beconstructed with Met-Ala- at the N-terminus so that upon expression theMet is cleaved off leaving Ala at the N-terminus. These mutant hIL-3polypeptides may also be expressed in E. coli by fusing a signal peptideto the N-terminus. This signal peptide is cleaved from the polypeptideas part of the secretion process. Secretion in E. coli can be used toobtain the correct amino acid at the N-terminus (e.g., Asn¹⁵ in the(15-125) hIL-3 polypeptide) due to the precise nature of the signalpeptidase. This is in contrast to the heterogeneity often observed atthe N-terminus of proteins expressed in the cytoplasm in E. coli.

[0713] The hIL-3 mutant polypeptides of the present invention may havehIL-3 or hIL-3-like activity. For example, they may possess one or moreof the biological activities of native hIL-3 and may be useful instimulating the production of hematopoietic cells by human or primateprogenitor cells. The hIL-3 muteins of the present invention andpharmaceutical compositions containing them may be useful in thetreatment of conditions in which hematopoietic cell populations havebeen reduced or destroyed due to disease or to treatments such asradiation or chemotherapy.

[0714] hIL-3 muteins of the present invention may also be useful asantagonists which block the hIL-3 receptor by binding specifically to itand preventing binding of the agonist.

[0715] One potential advantage of the (15-125) hIL-3 muteins of thepresent invention, particularly those which retain activity similar toor better than that of native hIL-3, is that it may be possible to use asmaller amount of the biologically active mutein to produce the desiredtherapeutic effect. This may make it possible to reduce the number oftreatments necessary to produce the desired therapeutic effect. The useof smaller amounts may also reduce the possibility of any potentialantigenic effects or other possible undesirable side effects. Forexample, if a desired therapeutic effect can be achieved with a smalleramount of polypeptide it may be possible to reduce or eliminate sideeffects associated with the administration of native IL-3 such as thestimulation of leukotriene and/or histamine release. The hIL-3 muteinsof the present invention may also be useful in the activation of stemcells or progenitors which have low receptor numbers. Pharmaceuticalcompositions containing (15-125) hIL-3 muteins of the present inventioncan be administered parenterally, intravenously, or subcutaneously.

[0716] As another aspect of the present invention, there is provided anovel method for producing the novel family of human IL-3 muteins. Themethod of the present invention involves culturing a suitable cell orcell line, which has been transformed with a vector containing a DNAsequence coding for expression of a novel hIL-3 mutant polypeptide.Suitable cells or cell lines may be bacterial cells. For example, thevarious strains of E. coli are well-known as host cells in the field ofbiotechnology. Examples of such strains include E. coli strains JM101[Yanish-Perron, et al. (1985)] and MON105 [Obukowicz, et al. (1992)].Various strains of B. subtilis may also be employed in this method. Manystrains of yeast cells known to those skilled in the art are alsoavailable as host cells for expression of the polypeptides of thepresent invention.

[0717] Also suitable for use in the present invention are mammaliancells, such as Chinese hamster ovary cells (CHO). General methods forexpression of foreign genes in mammalian cells are reviewed in: Kaufman,R. J. (1987) High level production of proteins in mammalian cells, inGenetic Engineering, Principles and Methods, Vol. 9, J. K. Setlow,editor, Plenum Press, New York. An expression vector is constructed inwhich a strong promoter capable of functioning in mammalian cells drivestranscription of a eukaryotic secretion signal peptide coding region,which is translationally fused to the coding region for the hIL-3variant. For example, plasmids such as pcDNA I/Neo, pRc/RSV, and pRc/CMV(obtained from Invitrogen Corp., San Diego, Calif.) can be used. Theeukaryotic secretion signal peptide coding region can be from the hIL-3gene itself or it can be from another secreted mammalian protein (Bayne,M. L. et al. (1987) Proc. Natl. Acad. Sci. USA 84, 2638-2642). Afterconstruction of the vector containing the hIL-3 variant gene, the vectorDNA is transfected into mammalian cells. Such cells can be, for example,the COS7, HeLa, BHK, CHO, or mouse L lines. The cells can be cultured,for example, in DMEM media (JRH Scientific). The hIL-3 variant secretedinto the media can be recovered by standard biochemical approachesfollowing transient expression 24-72 hours after transfection of thecells or after establishment of stable cell lines following selectionfor neomycin resistance. The selection of suitable mammalian host cellsand methods for transformation, culture, amplification, screening andproduct production and purification are known in the art. See, e.g.,Gething and Sambrook, Nature, 293:620-625 (1981), or alternatively,Kaufman et al, Mol. Cell. Biol., 5(7):1750-1759 (1985) or Howley et al.,U.S. Pat. No. 4,419,446. Another suitable mammalian cell line is themonkey COS-1 cell line. A similarly useful mammalian cell line is theCV-1 cell line.

[0718] Where desired, insect cells may be utilized as host cells in themethod of the present invention. See, e.g. Miller et al, GeneticEngineering, 8:277-298 (Plenum Press 1986) and references cited therein.In addition, general methods for expression of foreign genes in insectcells using Baculovirus vectors are described in: Summers, M. D. andSmith, G. E. (1987)—A manual of methods for Baculovirus vectors andinsect cell culture procedures, Texas Agricultural Experiment StationBulletin No. 1555. An expression vector is constructed comprising aBaculovirus transfer vector, in which a strong Baculovirus promoter(such as the polyhedron promoter) drives transcription of a eukaryoticsecretion signal peptide coding region, which is translationally fusedto the coding region for the hIL-3 variant polypeptide. For example, theplasmid pVL1392 (obtained from Invitrogen Corp., San Diego, Calif.) canbe used. After construction of the vector carrying the hIL-3 variantgene, two micrograms of this DNA is cotransfected with one microgram ofBaculovirus DNA (see Summers & Smith, 1987) into insect cells, strainSF9. Pure recombinant Baculovirus carrying the hIL-3 variant is used toinfect cells cultured, for example, in Excell 401 serum-free medium (JRHBiosciences, Lenexa, Kans.). The hIL-3 variant secreted into the mediumcan be recovered by standard biochemical approaches.

[0719] Another aspect of the present invention provides plasmid DNAvectors for use in the method of expression of these novel hIL-3muteins. These vectors contain the novel DNA sequences described abovewhich code for the novel polypeptides of the invention. Appropriatevectors which can transform microorganisms capable of expressing thehIL-3 muteins include expression vectors comprising nucleotide sequencescoding for the hIL-3 muteins joined to transcriptional and translationalregulatory sequences which are selected according to the host cellsused.

[0720] Vectors incorporating modified sequences as described above areincluded in the present invention and are useful in the production ofthe hIL-3 mutant polypeptides. The vector employed in the method alsocontains selected regulatory sequences in operative association with theDNA coding sequences of the invention and capable of directing thereplication and expression thereof in selected host cells.

[0721] Additional details may be found in co-filed U.S. PatentApplication Attorney docket No. 2713/1, which is hereby incorporated byreference in its entirety.

[0722] All references, patents or applications cited herein areincorporated by reference in their entirety.

[0723] The present invention also includes the construction andexpression of (15-125) human interleukin-3 muteins having four or moreamino acid substitutions in secretion vectors that optimize accumulationof correctly folded, active polypeptide. While many heterologousproteins have been secreted in E. coli there is still a great deal ofunpredictability and limited success (Stader and Silhavy 1990).Full-length hIL-3 is such a protein, where attempts to secrete theprotein in E. coli resulted in low secretion levels. Secretion of thevariant (15-125) hIL-3 mutant polypeptides of the present invention as afusion with a signal peptide such as LamB results in correctly foldedprotein that can be removed from the periplasm of E. coli by osmoticshock fractionation. This property of the variant (15-125) hIL-3 muteinsallows for the direct and rapid screening for bioactivity of thesecreted material in the crude osmotic shock fraction, which is asignificant advantage. Furthermore, it provides a means of using the(15-125) hIL-3 muteins to conduct structure activity relationship (SAR)studies of the hIL-3 molecule. A further advantage of secretion of(15-125) hIL-3 muteins fused to the LamB signal peptide is that thesecreted polypeptide has the correct N-terminal amino acid (Asn) due tothe precise nature of the cleavage of the signal peptide by signalpeptidase, as part of the secretion process.

[0724] The (15-125) hIL-3 muteins of the present invention may includehIL-3 polypeptides having Met-, Ala- or Met-Ala- attached to theN-terminus. When the muteins are expressed in the cytoplasm of E. coli,polypeptides with and without Met attached to the N-terminus areobtained. The methionine can in some cases be removed by methionineaminopeptidase.

[0725] Amino terminal sequences of hIL-3 muteins made in E. coli weredetermined using the method described by Hunkapillar et al., (1983). Itwas found that hIL-3 proteins made in E. coli from genes encodingMet-(15-125) hIL-3 were isolated as Met-(15-125) hIL-3. Proteinsproduced from genes encoding Met-Ala-(15-125) hIL-3 were produced asAla-(15-125) hIL-3. The N-termini of proteins made in the cytoplasm ofE. coli are affected by posttranslational processing by methionineaminopeptidase (Ben-Bassat et al., 1987) and possibly by otherpeptidases.

[0726] One method of creating the preferred hIL-3 (15-125) mutant genesis cassette mutagenesis [Wells, et al. (1985)] in which a portion of thecoding sequence of hIL-3 in a plasmid is replaced with syntheticoligonucleotides that encode the desired amino acid substitutions in aportion of the gene between two restriction sites. In a similar manneramino acid substitutions could be made in the full-length hIL-3 gene, orgenes encoding variants of hIL-3 in which from 1 to 14 amino acids havebeen deleted from the N-terminus and/or from 1 to 15 amino acids havebeen deleted from the C-terminus. When properly assembled theseoligonucleotides would encode hIL-3 variants with the desired amino acidsubstitutions and/or deletions from the N-terminus and/or C-terminus.These and other mutations could be created by those skilled in the artby other mutagenesis methods including; oligonucleotide-directedmutagenesis [Zoller and Smith (1982, 1983, 1984), Smith (1985), Kunkel(1985), Taylor, et al. (1985), Deng and Nickoloff (1992)] or polymerasechain reaction (PCR) techniques [Saiki, (1985)].

[0727] Pairs of complementary synthetic oligonucleotides encodingportions of the amino terminus of the hIL-3 gene can be made andannealed to each other. Such pairs would have protruding ends compatiblewith ligation to NcoI at one end. The NcoI site would include the codonfor the initiator methionine. At the other end of oligonucleotide pairs,the protruding (or blunt) ends would be compatible with a restrictionsite that occurs within the coding sequence of the hIL-3 gene. The DNAsequence of the oligonucleotide would encode sequence for amino acids ofhIL-3 with the exception of those substituted and/or deleted from thesequence.

[0728] The NcoI enzyme and the other restriction enzymes chosen shouldhave recognition sites that occur only once in the DNA of the plasmidchosen. Plasmid DNA can be treated with the chosen restrictionendonucleases then ligated to the annealed oligonucleotides. The ligatedmixtures can be used to transform competent JM101 cells to resistance toan appropriate antibiotic. Single colonies can be picked and the plasmidDNA examined by restriction analysis and/or DNA sequencing to identifyplasmids with mutant hIL-3 genes.

[0729] One example of a restriction enzyme which cleaves within thecoding sequence of the hIL-3 gene is ClaI whose recognition site is atcodons 20 and 21. The use of ClaI to cleave the sequence of hIL-3requires that the plasmid DNA be isolated from an E. coli strain thatfails to methylate adenines in the DNA at GATC recognition sites. Thisis because the recognition site for ClaI, ATCGAT, occurs within thesequence GATCGAT which occurs at codons 19, 20 and 21 in the hIL-3 gene.The A in the GATC sequence is methylated in most E. coli host cells.This methylation prevents ClaI from cleaving at that particularsequence. An example of a strain that does not methylate adenines isGM48.

[0730] Interpretation of Activity of Single Amino Acid Mutants in IL-3(15-125)

[0731] As illustrated in Tables 6 and 9, there are certain positions inthe IL-3 (15-125) molecule which are intolerant of substitutions, inthat most or all substitutions at these positions resulted in aconsiderable decrease in bioactivity. There are two likely classes ofsuch “down-mutations”: mutations that affect overall protein structure,and mutations that interfere directly with the interaction between theIL-3 molecule and its receptor. Mutations affecting thethree-dimensional structure of the protein will generally lie in theinterior of the protein, while mutations affecting receptor binding willgenerally lie on the surface of the protein. Although thethree-dimensional structure of IL-3 is unknown, there are simplealgorithms which can aid in the prediction of the structure. One suchalgorithm is the use of “helical wheels” (Kaiser, E. T. & Kezdy, F. J.,Science, 223:249-255 (1984)). In this method, the presence of alphahelical protein structures can be predicted by virtue of theiramphipathic nature. Helices in globular proteins commonly have anexposed hydrophilic side and a buried hydrophobic side. As a broadgeneralization, in globular proteins, hydrophobic residues are presentin the interior of the protein, and hydrophilic residues are present onthe surface. By displaying the amino acid sequence of a protein on sucha “helical wheel” it is possible to derive a model for which amino acidsin alpha helices are exposed and which are buried in the core of theprotein. Such an analysis of the IL-3 (15-125) molecule predicts thatthe following helical residues are buried in the core:

[0732] M19, I20, I23, I24, L27, L58, F61, A64, L68, A71, I74, I77, L78,L81, W104, F107, L111, Y114, L115, L118.

[0733] In addition, cysteine residues at positions 16 and 84 are linkedby a disulfide bond, which is important for the overall structure or“folding” of the protein. Finally, mutations which result in a majordisruption of the protein structure may be expressed at low level in thesecretion system used in our study, for a variety of reasons: eitherbecause the mis-folded protein is poorly recognized by the secretionmachinery of the cell; because mis-folding of the protein results inaggregation, and hence the protein cannot be readily extracted from thecells; or because the mis-folded protein is more susceptible todegradation by cellular proteases. Hence, a block in secretion mayindicate which positions in the IL-3 molecule which are important formaintenance of correct protein structure.

[0734] In order to retain the activity of a variant of IL-3, it isnecessary to retain both the structural integrity of the protein, andretain the specific residues important for receptor contact. Hence it ispossible to define specific amino acid residues in IL-3 (15-125) whichmust be retained in order to preserve biological activity.

[0735] Residues predicted to be important for interaction with thereceptor: D21, E22, E43, D44, L48, R54, R94, D103, K110, F113.

[0736] Residues predicted to be structurally important: C16, L58, F61,A64, I74, L78, L81, C84, P86, P92, P96, F107, L111, L115, L118.

[0737] The hIL-3 muteins of the present invention may be useful in thetreatment of diseases characterized by a decreased levels of eithermyeloid, erythroid, lymphoid, or megakaryocyte cells of thehematopoietic system or combinations thereof. In addition, they may beused to activate mature myeloid and/or lymphoid cells. Among conditionssusceptible to treatment with the polypeptides of the present inventionis leukopenia, a reduction in the number of circulating leukocytes(white cells) in the peripheral blood. Leukopenia may be induced byexposure to certain viruses or to radiation. It is often a side effectof various forms of cancer therapy, e.g., exposure to chemotherapeuticdrugs and of infection or hemorrhage. Therapeutic treatment ofleukopenia with these hIL-3 mutant polypeptides of the present inventionmay avoid undesirable side effects caused by treatment with presentlyavailable drugs.

[0738] The hIL-3 muteins of the present invention may be useful in thetreatment of neutropenia and, for example, in the treatment of suchconditions as aplastic anemia, cyclic neutropenia, idiopathicneutropenia, Chediak-Higashi syndrome, systemic lupus erythematosus(SLE), leukemia, myelodysplastic syndrome and myelofibrosis.

[0739] Many drugs may cause bone marrow suppression or hematopoieticdeficiencies. Examples of such drugs are AZT, DDI, alkylating agents andanti-metabolites used in chemotherapy, antibiotics such aschloramphenicol, penicillin and sulfa drugs, phenothiazones,tranquilizers such as meprobamate, and diuretics. The hIL-3 muteins ofthe present invention may be useful in preventing or treating the bonemarrow suppression or hematopoietic deficiencies which often occur inpatients treated with these drugs.

[0740] Hematopoietic deficiencies may also occur as a result of viral,microbial or parasitic infections and as a result of treatment for renaldisease or renal failure, e.g., dialysis. The hIL-3 muteins of thepresent invention may be useful in treating such hematopoieticdeficiency.

[0741] The treatment of hematopoietic deficiency may includeadministration of the hIL-3 mutein of a pharmaceutical compositioncontaining the hIL-3 mutein to a patient. The hIL-3 muteins of thepresent invention may also be useful for the activation andamplification of hematopoietic precursor cells by treating these cellsin vitro with the muteins of the present invention prior to injectingthe cells into a patient.

[0742] Various immunodeficiencies e.g., in T and/or B lymphocytes, orimmune disorders, e.g., rheumatoid arthritis, may also be beneficiallyaffected by treatment with the hIL-3 mutant polypeptides of the presentinvention. Immunodeficiencies may be the result of viral infections e.g.HTLVI, HTLVII, HTLVIII, severe exposure to radiation, cancer therapy orthe result of other medical treatment. The hIL-3 mutant polypeptides ofthe present invention may also be employed, alone or in combination withother hematopoietins, in the treatment of other blood cell deficiencies,including thrombocytopenia (platelet deficiency), or anemia. Other usesfor these novel polypeptides are in the treatment of patients recoveringfrom bone marrow transplants in vivo and ex vivo, and in the developmentof monoclonal and polyclonal antibodies generated by standard methodsfor diagnostic or therapeutic use.

[0743] Other aspects of the present invention are methods andtherapeutic compositions for treating the conditions referred to above.Such compositions comprise a therapeutically effective amount of one ormore of the hIL-3 muteins of the present invention in a mixture with apharmaceutically acceptable carrier. This composition can beadministered either parenterally, intravenously or subcutaneously. Whenadministered, the therapeutic composition for use in this invention ispreferably in the form of a pyrogen-free, parenterally acceptableaqueous solution. The preparation of such a parenterally acceptableprotein solution, having due regard to pH, isotonicity, stability andthe like, is within the skill of the art.

[0744] The dosage regimen involved in a method for treating theabove-described conditions will be determined by the attending physicianconsidering various factors which modify the action of drugs, e.g. thecondition, body weight, sex and diet of the patient, the severity of anyinfection, time of administration and other clinical factors. Generally,a daily regimen may be in the range of 0.2-150 μg/kg of non-glycosylatedIL-3 protein per kilogram of body weight. This dosage regimen isreferenced to a standard level of biological activity which recognizesthat native IL-3 generally possesses an EC₅₀ at or about 10 picoMolar to100 picoMolar in the AML proliferation assay described herein.Therefore, dosages would be adjusted relative to the activity of a givenmutein vs. the activity of native (reference) IL-3 and it would not beunreasonable to note that dosage regimens may include doses as low as0.1 microgram and as high as 1 milligram per kilogram of body weight perday. In addition, there may exist specific circumstances where dosagesof TL-3 mutein would be adjusted higher or lower than the range of10-200 micrograms per kilogram of body weight. These includeco-administration with other CSF or growth factors; co-administrationwith chemotherapeutic drugs and/or radiation; the use of glycosylatedIL-3 mutein; and various patient-related issues mentioned earlier inthis section. As indicated above, the therapeutic method andcompositions may also include co-administration with other humanfactors. A non-exclusive list of other appropriate hematopoietins, CSFsand interleukins for simultaneous or serial co-administration with thepolypeptides of the present invention includes GM-CSF, CSF-1, G-CSF,Meg-CSF, M-CSF, erythropoietin (EPO), IL-1, IL-4, IL-2, IL-5, IL-6,IL-7, IL-8, IL-9, IL-10, IL-11, LIF, B-cell growth factor, B-celldifferentiation factor and eosinophil differentiation factor, stem cellfactor (SCF) also known as steel factor or c-kit ligand, or combinationsthereof. The dosage recited above would be adjusted to compensate forsuch additional components in the therapeutic composition. Progress ofthe treated patient can be monitored by periodic assessment of thehematological profile, e.g., differential cell count and the like.

[0745] Materials and Methods for hIL-3 Mutein Expression in E. coli

[0746] Unless noted otherwise, all specialty chemicals were obtainedfrom Sigma Co., (St. Louis, Mo.). Restriction endonucleases, T4poly-nucleotides kinase, E. coli DNA polymerase T large fragment(Klenow) and T4 DNA ligase were obtained from New England Biolabs(Beverly, Mass.).

[0747]Escherichia coli Strains

[0748] Strain JM101: delta (pro lac), supE, thi, F′(traD36, rpoAB,lacI-Q, lacZdeltaM15) (Messing, 1979). This strain can be obtained fromthe American Type Culture Collection (ATCC), 12301 Parklawn Drive,Rockville, Md. 20852, accession number 33876. MON 105 (W3110 rpoH358) isa derivative of W3110 (Bachmann, 1972) and has been assigned ATCCaccession number 55204. Strain GM48: dam-3, dcm-6, gal, ara, lac, thr,leu, tonA, tsx (Marinus, 1973) was used to make plasmid DNA that is notmethylated at the sequence GATC.

[0749] Genes and Plasmids

[0750] The gene used for hIL-3 production in E. coli was obtained fromBritish Biotechnology Incorporated, Cambridge, England, catalogue numberBBG14. This gene is carried on a pUC based plasmid designated pP0518.

[0751] The plasmids used for production of hIL-3 in E. coli containgenetic elements whose use has been described (Olins et al., 1988; Olinsand Rangwala, 1990). The replicon used is that of pBR327 (Covarrubias,et al., 1981) which is maintained at a copy number of about 100 in thecell (Soberon et al., 1980). A gene encoding the beta-lactamase proteinis present on the plasmids. This protein confers ampicillin resistanceon the cell. This resistance serves as a selectable phenotype for thepresence of the plasmid in the cell.

[0752] For cytoplasmic expression vectors the transcription promoter wasderived from the recA gene of E. coli (Sancar et al., 1980). Thispromoter, designated precA, includes the RNA polymerase binding site andthe lexA repressor binding site (the operator). This segment of DNAprovides high level transcription that is regulated even when the recApromoter is on a plasmid with the pBR327 origin of replication (Olins etal., 1988) incorporated herein by reference.

[0753] In secretion expression plasmids the transcription promoter wasderived from the ara B, A, and D genes of E. coli (Greenfield et al.,1978). This promoter is designated pAraBAD and is contained on a 323base pair SacII, BglII restriction fragment. The LamB secretion leader(Wong et al., 1988, Clement et al., 1981) was fused to the N-terminus ofthe hIL-3 gene at the recognition sequence for the enzyme NcoI(5′CCATGG3′). The hIL-3 genes used were engineered to have a HindIIIrecognition site (5′AAGCTT3′) following the coding sequence of the gene.

[0754] These hIL-3 variants were expressed as a fusion with the LamBsignal peptide shown in FIG. 8, operatively joined to the araBADpromoter (Greenfield, 1978) and the g10-L ribosome binding site (Olinset al. 1988). The processed form was selectively released from theperiplasm by osmotic shock as a correctly folded and fully activemolecule. Secretion of (15-125) hIL-3 was further optimized by using lowinducer (arabinose) concentration and by growth at 30° C. Theseconditions resulted in lower accumulation levels of unprocessed LamBsignal peptide (15-125) hIL-3 fusion, maximal accumulation levels ofprocessed (15-125) hIL-3 and selective release of (15-125) hIL-3 byosmotic shock fractionation. The use of a tightly regulated promotersuch as araBAD from which the transcription level and hence theexpression level can be modulated allowed for the optimization ofsecretion of (15-125) hIL-3.

[0755] The ribosome binding site used is that from gene 10 of phage T7(Olins et al., 1988). This is encoded in a 100 base pair (bp) fragmentplaced adjacent to precA. In the plasmids used herein, the recognitionsequence for the enzyme NcoI (CCATGG) follows the g10-L. It is at thisNcoI site that the hIL-3 genes are joined to the plasmid. It is expectedthat the nucleotide sequence at this junction will be recognized in mRNAas a functional start site for translation (Olins et al., 1988). ThehIL-3 genes used were engineered to have a HindIII recognition site(AAGCTT) downstream from the coding sequence of the gene. At thisHindIII site is a 514 base pair RsaI fragment containing the origin ofreplication of the single stranded phage f1 (Dente et al., 1983; Olins,et al., 1990) both incorporated herein by reference. A plasmidcontaining these elements is pMON2341. Another plasmid containing theseelements is pMON5847 which has been deposited at the American TypeCulture Collection, 12301 Parklawn Drive, Rockville, Md. 20852 under theaccession number ATCC 68912.

[0756] Synthesis of Oligonucleotides

[0757] Oligonucleotides were synthesized on Nucleotide Synthesizer model380A or 380B from Applied Biosystems, Inc. (Foster City, Calif.).Oligonucleotides were purified by polyacrylamide gel electrophoresis atconcentrations from 12-20% (19:1 crosslinked) in 0.5×Tris borate buffer(0.045 M Tris, 0.045 M boric acid, 1.25 mM EDTA) followed by passagethrough a Nensorb column obtained from New England Nuclear (Boston,Mass.) using a PREP Automated Sample Processor obtained from DuPont, Co.(Wilmington, Del.).

[0758] Quantitation of Synthetic Oligonucleotides

[0759] Synthetic oligonucleotides were resuspended in water andquantitated by reading the absorbance at 260 nm on a Beckman DU40Spectrophotometer (Irvine, Calif.) using a one centimeter by onemillimeter quartz cuvette (Maniatis, 1982). The concentration wasdetermined using an extinction coefficient of 1×10⁴ (Voet et al., 1963;Mahler and Cordes, 1966). The oligonucleotides were then diluted to adesired concentration.

[0760] Quantitation of synthetic DNA fragments can also be achieved byadding 10 to 100 picomoles of DNA to a solution containing kinase buffer(25 mM Tris pH 8.0, 10 mM MgCl₂, 10 mM DTT and 2 mM spermidine). To thereaction mix is added ATP to 20 micromolar, ATP radiolabeled at thegamma phosphate (5000-10,0000 dpm/pmol) and 5 units of T4 polynucleotidekinase. Radiolabelled material is obtained from New England Nuclear(Boston, Mass.). The 10 microliter mixture is incubated at 37° C. forone hour. A 1 microliter aliquot of the mixture was chromatographed onDEAE paper (Whatman) in 0.3 M ammonium bicarbonate. The counts thatremained at the origin were used to determine the concentration of thesynthetic DNA.

[0761] Recombinant DNA Methods

[0762] Isolation of plasmid DNA from E. coli cultures was performed asdescribed (Birnboim and Doly, 1979). Some DNAs were purified by Magic™columns, available from Promega (Madison, Wis.).

[0763] Purified plasmid DNA was treated with restriction endonucleasesaccording to manufacturer's instructions. Analysis of the DNA fragmentsproduced by treatment with restriction enzymes was done by agarose orpolyacrylamide gel electrophoresis. Agarose (DNA grade from Fisher,Pittsburgh Pa.) was used at a concentration of 1.0% in a Tris-acetaterunning buffer (0.04 M Tris-acetate, 0.001M EDTA). Polyacrylamide(BioRad, Richmond Calif.) was used at a concentration of 6% (19:1crosslinked) in 0.5×Tris-borate buffer (0.045 M Tris, 0.045 M boricacid, 1.25 mM EDTA), hereafter referred to as PAGE.

[0764] DNA polymerase I, large fragment, Klenow enzyme was usedaccording to manufacturers instructions to catalyze the addition ofmononucleotides from 5′ to 3′ of DNA fragments which had been treatedwith restriction enzymes that leave protruding ends. The reactions wereincubated at 65° C. for 10 minutes to heat inactivate the Klenow enzyme.

[0765] The synthetic oligonucleotides were made without 5′ or 3′terminal phosphates. In cases where such oligonucleotides were ligatedend to end, the oligonucleotides were treated at a concentration of 10picomoles per microliter with T4 polynucleotide kinase in the followingbuffer: 25 mM Tris, pH 8.0, 10 mM MgCl₂, 10 mM dithiothreitol, 2 mMspermidine, 1 mM rATP. After incubation for 30 minutes at 37° C., thesamples were incubated at 65° C. for five minutes to heat inactivate thekinase.

[0766] Synthetic Gene Assembly

[0767] The (15-125) hIL-3 gene was divided into four regions separatedby five convenient restriction sites. In each of the four regionssynthetic oligonucleotides were designed so that they would anneal incomplementary pairs, with protruding single stranded ends, and when thepairs were properly assembled would result in a DNA sequence thatencoded a portion of the hIL-3 gene. Amino acid substitutions in thehIL-3 gene were made by designing the oligonucleotides to encode thedesired substitutions. The complementary oligonucleotides were annealedat concentration of 1 picomole per microliter in ligation buffer plus 50mM NaCl. The samples were heated in a 100 ml beaker of boiling water andpermitted to cool slowly to room temperature. One picomole of each ofthe annealed pairs of oligonucleotides were ligated with approximately0.2 picomoles of plasmid DNA, digested with the appropriate restrictionenzymes, in ligation buffer (25 mM Tris pH 8.0, 10 mM MgCl₂, 10 mMdithiothreitol, 1 mM ATP, 2 mM spermidine) with T4 DNA ligase obtainedfrom New England Biolabs (Beverly, Mass.) in a total volume of 20 μl atroom temperature overnight.

[0768] DNA fragments were isolated from agarose gels by intercepting therestriction fragments on DEAE membranes from Schleicher and Schuell(Keene, N.H.) and eluting the DNA in 10 mM Tris, 1 mM EDTA, 1 M NaCl at55° C. for 1 hour, according to manufacturer's directions. The solutionscontaining the DNA fragment were concentrated and desalted by usingCentricon 30 concentrators from Amicon (W. R. Grace, Beverly Mass.)according to the manufacturer's directions. Ligations were performed at15° C. overnight, except as noted, in ligation buffer.

[0769] Polymerase Chain Reaction

[0770] Polymerase Chain Reaction (hereafter referred to as PCR)techniques (Saiki, 1985) used the reagent kit and thermal cycler fromPerkin-Elmer Cetus (Norwalk, Conn.). PCR is based on a thermostable DNApolymerase from Thermus aquaticus. The PCR technique is a DNAamplification method that mimics the natural DNA replication process inthat the number of DNA molecules doubles after each cycle, in a waysimilar to in vivo replication. The DNA polymerase mediated extension isin a 5′ to 3′ direction. The term “primer” as used herein refers to anoligonucleotide sequence that provides an end to which the DNApolymerase can add nucleotides that are complementary to a nucleotidesequence. The latter nucleotide sequence is referred to as the“template”, to which the primers are annealed. The amplified PCR productis defined as the region comprised between the 5′ ends of the extensionprimers. Since the primers have defined sequences, the product will havediscrete ends, corresponding to the primer sequences. The primerextension reaction was carried out using 20 picomoles (pmoles) of eachof the oligonucleotides and 1 picogram of template plasmid DNA for 35cycles (1 cycle is defined as 94 degrees C. for one minute, 50 degreesC. for two minutes and 72 degrees for three minutes.). The reactionmixture was extracted with an equal volume of phenol/chloroform (50%phenol and 50% chloroform, volume to volume) to remove proteins. Theaqueous phase, containing the amplified DNA, and solvent phase wereseparated by centrifugation for 5 minutes in a microcentrifuge (Model5414 Eppendorf Inc, Fremont Calif.). To precipitate the amplified DNAthe aqueous phase was removed and transferred to a fresh tube to whichwas added {fraction (1/10)} volume of 3M NaOAc (pH 5.2) and 2.5 volumesof ethanol (100% stored at minus 20 degrees C.). The solution was mixedand placed on dry ice for 20 minutes. The DNA was pelleted bycentrifugation for 10 minutes in a microcentrifuge and the solution wasremoved from the pellet. The DNA pellet was washed with 70% ethanol,ethanol removed and dried in a speedvac concentrator (Savant,Farmingdale, N.Y.). The pellet was resuspended in 25 microliters of TE(20 mM Tris-HCl pH 7.9, 1 mM EDTA). Alternatively the DNA wasprecipitated by adding equal volume of 4M NH₄OAc and one volume ofisopropanol [Treco et al., (1988)]. The solution was mixed and incubatedat room temperature for 10 minutes and centrifuged. These conditionsselectively precipitate DNA fragments larger than 20 bases and were usedto remove oligonucleotide primers. One quarter of the reaction wasdigested with restriction enzymes [Higuchi, (1989)] an on completionheated to 70 degrees C. to inactivate the enzymes.

[0771] Recovery of Recombinant Plasmids from Ligation Mixes

[0772]E. coli JM101 cells were made competent to take up DNA. Typically,20 to 100 ml of cells were grown in LB medium to a density ofapproximately 150 Klett units and then collected by centrifugation. Thecells were resuspended in one half culture volume of 50 mM CaCl₂ andheld at 4° C. for one hour. The cells were again collected bycentrifugation and resuspended in one tenth culture volume of 50 mMCaCl₂. DNA was added to a 150 microliter volume of these cells, and thesamples were held at 4° C. for 30 minutes. The samples were shifted to42° C. for one minute, one milliliter of LB was added, and the sampleswere shaken at 37° C. for one hour. Cells from these samples were spreadon plates containing ampicillin to select for transformants. The plateswere incubated overnight at 37° C. Single colonies were picked, grown inLB supplemented with ampicillin overnight at 37° C. with shaking. Fromthese cultures DNA was isolated for restriction analysis.

[0773] Culture Medium

[0774] LB medium (Maniatis et al., 1982) was used for growth of cellsfor DNA isolation. M9 minimal medium supplemented with 1.0% casaminoacids, acid hydrolyzed casein, Difco (Detroit, Mich.) was used forcultures in which recombinant hIL-3 was produced. The ingredients in theM9 medium were as follows: 3 g/liter KH₂PO₄, 6 g/l Na₂HPO₄, 0.5 g/lNaCl, 1 g/l NH₄Cl, 1.2 mM MgSO₄, 0.025 mM CaCl₂, 0.2% glucose (0.2%glycerol with the AraBAD promoter), 1% casamino acids, 0.1 ml/l traceminerals (per liter 108 g FeCl₃.6H₂O, 4.0 g ZnSO₄.7H₂O, 7.0 CoCl₂.2H₂O,7.0 g Na₂MoO₄.2H₂O, 8.0 g CuSO₄.5H₂O, 2.0 g H₃BO₃, 5.0 g MnSO₄.H₂O, 100ml concentrated HCl). Bacto agar was used for solid media and ampicillinwas added to both liquid and solid LB media at 200 micrograms permilliliter.

[0775] DNA Sequence Analysis

[0776] The nucleotide sequencing of plasmid DNA was determined using aGenesis 2000 sequencer obtained from DuPont (Wilmington, Del.) accordingto the methods of Prober et al. (1987) and Sanger et al. (1977). SomeDNA sequences were performed using Sequenase™ polymerase (U.S.Biochemicals, Cleveland, Ohio) according to manufacturer's directions.

[0777] Production of Recombinant hIL-3 Muteins in E. coli with VectorsEmploying the recA Promoter

[0778]E. coli strains harboring the plasmids of interest were grown at37° C. in M9 plus casamino acids medium with shaking in a Gyrotory waterbath Model G76 from New Brunswick Scientific (Edison, N.J.). Growth wasmonitored with a Klett Summerson meter (green 54 filter), Klett Mfg. Co.(New York, N.Y.). At a Klett value of approximately 150, an aliquot ofthe culture (usually one milliliter) was removed for protein analysis.To the remaining culture, nalidixic acid (10 mg/ml) in 0.1 N NaOH wasadded to a final concentration of 50 μg/ml. The cultures were shaken at37° C. for three to four hours after addition of nalidixic acid. A highdegree of aeration was maintained throughout the bacterial growth inorder to achieve maximal production of the desired gene product. Thecells were examined under a light microscope for the presence ofrefractile bodies (RBs). One milliliter aliquots of the culture wereremoved for analysis of protein content.

[0779] Production of Recombinant hIL-3 Proteins from the pAraBADPromoter in E. coli

[0780]E. coli strains harboring the plasmids of interest were grown at30° C. with shaking in M9 medium plus casamino acids and glycerol.Growth was monitored with a Klett Summerson calorimeter, using a green54 filter. At a Klett value of about 150, an aliquot of the culture(usually one milliliter) was removed for protein analysis. To theremaining culture, 20% arabinose was added to a final concentration of0.05%. The cultures were shaken at 30° C. for three to four hours afteraddition of arabinose. A high degree of aeration was maintainedthroughout the bacterial growth in order to achieve maximal productionof the desired gene product. One milliliter aliquots of the culture wereremoved for analysis of protein content.

[0781] Secretion and Osmotic Shock

[0782] Three hour post induction samples were fractionated by osmoticshock [Neu and Heppel (1965)]. The optical density (Klett value) of thecultures was determined and 1 ml of cells were centrifuged in a Sigmamicrocentrifuge (West Germany) model 202MK in 1.5 mls snap topmicrocentrifuge tubes for 5 minutes at 10,000 rpm. The cell pellet wasresuspended very gently by pipeting in a room temperature sucrosesolution (20% sucrose w/v, 30 mM Tris-Hcl pH7.5, 1 mM EDTA), using 1μl/1 Klett unit. Following a 10 minute incubation at room temperature,the cells were centrifuged for 5 minutes at 10,000 rpm. The sucrosefraction was carefully removed from the cell pellet. The cell pellet wasthen resuspended very gently by pipeting in ice cold distilled water,using 1 μl/l Klett unit. Following a 10 minute incubation on ice, thecells were centrifuged for 5 minutes at 12,000 rpm. The water fractionwas carefully removed. Equal volumes of the sucrose and water fractionswere pooled and aliquoted to provide samples for activity screening.

[0783] Analysis of Protein Content of E. coli Cultures Producing hIL-3Mutant Polypeptides

[0784] Bacterial cells from cultures treated as described above werecollected from the medium by centrifugation. Aliquots of these cellswere resuspended in SDS loading buffer (4×: 6 g SDS, 10 mlbeta-mercaptoethanol, 25 ml upper Tris gel stock (0.5 M Tris HCl pH 6.8,0.4% SDS) brought to 50 ml with glycerol, 0.2% bromophenol blue wasadded) at a concentration of one microliter per Klett unit. Thesesamples were incubated at 85° C. for five minutes and vortexed. Five orten microliter aliquots of these samples were loaded on 15%polyacrylamide gels prepared according to the method of Laemmli (1970).Protein bands were visualized by staining the gels with a solution ofacetic acid, methanol and water at 5:1:5 ratio (volume to volume) towhich Coomassie blue had been added to a final concentration of 1%.After staining, the gels were washed in the same solution without theCoomassie blue and then washed with a solution of 7% acetic acid, 5%methanol. Gels were dried on a gel drier Model SE1160 obtained fromHoeffer (San Francisco, Calif.). The amount of stained protein wasmeasured using a densitometer obtained from Joyce-Loebl (Gateshead,England). The values obtained were a measure of the amount of thestained hIL-3 protein compared to the total of the stained protein ofthe bacterial cells.

[0785] Western Blot Analysis of hIL-3 Muteins made in E. coli

[0786] In some E. coli cultures producing hIL-3, the level ofaccumulation of the hIL-3 protein is lower than 5% of total bacterialprotein. To detect hIL-3 produced at this level, Western blot analysiswas used. Proteins from cultures induced with nalidixic acid orarabinose were run on polyacrylamide gels as described above except thatvolumes of sample loaded were adjusted to produce appropriate signals.After electrophoresis, the proteins were electroblotted to APT paper,Transa-bind, Schleicher and Schuell (Keene, N.H.) according to themethod of Renart et al. (1979). Antisera used to probe these blots hadbeen raised in rabbits, using peptides of the sequence of amino acids 20to 41 and 94 to 118 of hIL-3 as the immunogens. The presence of boundantibody was detected with Staphylococcal protein A radiolabeled with¹²⁵I, obtained from New England Nuclear (Boston, Mass.).

[0787] Fractionation of E. coli Cells Producing hIL-3 Proteins in theCytoplasm

[0788] Cells from E. coli cultures harboring plasmids that produce hIL-3muteins were induced with nalidixic acid. After three hours, the hIL-3muteins accumulated in retractile bodies. The first step in purificationof the hIL-3 muteins was to sonicate cells. Aliquots of the culture wereresuspended from cell pellets in sonication buffer: 10 mM Tris, pH 8.0,1 mM EDTA, 50 mM NaCl and 0.1 mM PMSF. These resuspended cells weresubjected to several repeated sonication bursts using the microtip froma Sonicator cell disrupter, Model W-375 obtained from HeatSystems-Ultrasonics Inc. (Farmingdale, N.Y.). The extent of sonicationwas monitored by examining the homogenates under a light microscope.When nearly all of the cells had been broken, the homogenates werefractionated by centrifugation. The pellets, which contain most of therefractile bodies, are highly enriched for hIL-3 muteins.

[0789] Methods: Extraction, Refolding and Purification of Interleukin-3(IL-3) Muteins Expressed as Refractile Bodies in E. coli.

[0790] Extraction of refractile bodies (RB's):

[0791] For each gram of RB's (and typically one gram is obtained from a300 ml E. coli culture), 5 ml of a solution containing 6M guanidinehydrochloride (GnHCl), 50 mM 2-N-cyclohexylaminoethanesulfonic acid(CHES) pH 9.5 and 20 mM dithiothreitol (DTT) was added. The RB's wereextracted with a Bio-Homogenizer for 15-30 seconds and gently rocked for2 hours at 5 degrees centigrade (5° C.) to allow the protein tocompletely reduce and denature.

[0792] Refolding of the IL-3 Muteins

[0793] The protein solution was transferred to dialysis tubing (1000molecular weight cut-off) and dialyzed against at least 100 volumes of4M GnHCl-50 mM CHES pH 8.0. The dialysis was continued overnight at 5°C. while gently stirring. Subsequently dialysis was continued against atleast 100 volumes of 2M GnHCl-50 mM CHES pH 8.0 and dialyzed overnightat 5° C. while gently stirring.

[0794] Purification of the IL-3 Muteins

[0795] The protein solution was removed from the dialysis tubing andacidified by the addition of 40% acetonitrile (CH₃CN)-0.2%trifluoroacetic acid (TFA) to a final concentration of 20% CH₃CN-0.1%TFA. This was centrifuged (16,000×g for 5 minutes) to clarify and thesupernatant was loaded onto a Vydac C-18 reversed phase column (10×250mm) available from Vydac (Hesperia, Calif.) previously equilibrated in20% CH₃CN-0.1% TFA. The column was eluted with a linear gradient (0.2%CH₃CN/minute) between 40-50% CH₃CN-0.1% TFA at a flow rate of 3ml/minute while collecting 1.5 ml fractions. The fractions were analyzedby polyacrylamide gel electrophoresis (SDS-PAGE) and the appropriatefractions pooled. The pooled material was dried by lyophilization or ina Speed Vac concentrator. The dry powder was reconstituted with 10 mMammonium bicarbonate pH 7.5, centrifuged (16,000×g for 5 minutes) toclarify and assayed for protein concentration by the method of Bradford(1976) with bovine serum albumin as the standard. Such protein can befurther analyzed by additional techniques such as, SDS-PAGE,electrospray mass spectrometry, reverse phase HPLC, capillary zoneelectrophoresis, amino acid composition analysis, and ELISA(enzyme-linked immunosorbent assay).

[0796] hIL-3 SANDWICH ELISA

[0797] IL-3 protein concentrations can be determined using a sandwichELISA based on an affinity purified polyclonal goat anti-rhIL-3.Microtiter plates (Dynatech Immulon II) were coated with 150 μlgoat-anti-rhIL-3 at a concentration of approximately 1 μg/ml in 100 mMNaHCO3, pH 8.2. Plates were incubated overnight at room temperature in achamber maintaining 100% humidity. Wells were emptied and the remainingreactive sites on the plate were blocked with 200 μl of solutioncontaining 10 mM PBS, 3% BSA and 0.05% Tween 20, pH 7.4 for 1 hour at37° C. and 100% humidity. Wells were emptied and washed 4× with 150 mMNaCl containing 0.05% Tween 20 (wash buffer). Each well then received150 μl of dilution buffer (10 mM PBS containing 0.1% BSA, 0.01% Tween20, pH 7.4), containing rhIL-3 standard, control, sample or dilutionbuffer alone. A standard curve was prepared with concentrations rangingfrom 0.125 ng/ml to 5 ng/ml using a stock solution of rhIL-3(concentration determined by amino acid composition analysis). Plateswere incubated 2.5 hours at 37° C. and 100% humidity. Wells were emptiedand each plate was washed 4× with wash buffer. Each well then received150 μl of an optimal dilution (as determined in a checkerboard assayformat) of goat anti-rhIL-3 conjugated to horseradish peroxidase. Plateswere incubated 1.5 hours at 37° C. and 100% humidity. Wells were emptiedand each plate was washed 4× with wash buffer. Each well then received150 ul of ABTS substrate solution (Kirkegaard and Perry). Plates wereincubated at room temperature until the color of the standard wellscontaining 5 ng/ml rhIL-3 had developed enough to yield an absorbancebetween 0.5-1.0 when read at a test wavelength of 410 nm and a referencewavelength of 570 nm on a Dynatech microtiter plate reader.Concentrations of immunoreactive rhIL-3 in unknown samples werecalculated from the standard curve using software supplied with theplate reader.

[0798] AML Proliferation Assay for Bioactive Human Interleukin-3

[0799] The factor-dependent cell line AML 193 was obtained from theAmerican Type Culture Collection (ATCC, Rockville, Md.). This cell line,established from a patient with acute myelogenous leukemia, is a growthfactor dependent cell line which displayed enhanced growth in GM/CSFsupplemented medium (Lange, B., et al., (1987); Valtieri, M., et al.,(1987). The ability of AML 193 cells to proliferate in the presence ofhuman IL-3 has also been documented. (Santoli, D., et al., (1987)). Acell line variant was used, AML 193 1.3, which was adapted for long termgrowth in IL-3 by washing out the growth factors and starving thecytokine dependent AML 193 cells for growth factors for 24 hours. Thecells were then replated at 1×10⁵ cells/well in a 24 well plate in mediacontaining 100 U/ml IL-3. It took approximately 2 months for the cellsto grow rapidly in IL-3. These cells were maintained as AML 193 1.3thereafter by supplementing tissue culture medium (see below) with humanIL-3.

[0800] AML 193 1.3 cells were washed 6 times in cold Hanks balanced saltsolution (HBSS, Gibco, Grand Island, N.Y.) by centrifuging cellsuspensions at 250×g for 10 minutes followed by decantation ofsupernatant. Pelleted cells were resuspended in HBSS and the procedurewas repeated until six wash cycles were completed. Cells washed sixtimes by this procedure were resuspended in tissue culture medium at adensity ranging from 2×10⁵ to 5×10⁵ viable cells/ml. This medium wasprepared by supplementing Iscove's modified Dulbecco's Medium (TMDM,Hazleton, Lenexa, Kans.) with albumin, transferrin, lipids and2-mercaptoethanol. Bovine albumin (Boehringer-Mannheim, Indianapolis,Ind.) was added at 500 μg/ml; human transferrin (Boehringer-Mannheim,Indianapolis, Ind.) was added at 100 μg/ml; soybean lipid(Boehringer-Mannheim, Indianapolis, Ind.) was added at 50 μg/ml; and2-mercaptoethanol (Sigma, St. Louis, Mo.) was added at 5×10⁻⁵ M.

[0801] Serial dilutions of human interleukin-3 or human interleukin-3variant protein (hIL-3 mutein) were made in triplicate series in tissueculture medium supplemented as stated above in 96 well Costar 3596tissue culture plates. Each well contained 50 μl of medium containinginterleukin-3 or interleukin-3 variant protein once serial dilutionswere completed. Control wells contained tissue culture medium alone(negative control). AML 193 1.3 cell suspensions prepared as above wereadded to each well by pipetting 50 μl (2.5×10⁴ cells) into each well.Tissue culture plates were incubated at 37° C. with 5% CO₂ in humidifiedair for 3 days. On day 3, 0.5 μCi ³H-thymidine (2 Ci/mM, New EnglandNuclear, Boston, Mass.) was added in 50 μl of tissue culture medium.Cultures were incubated at 37° C. with 5% CO₂ in humidified air for18-24 hours. Cellular DNA was harvested onto glass filter mats(Pharmacia LKB, Gaithersburg, Md.) using a TOMTEC cell harvester(TOMTEC, Orange, Conn.) which utilized a water wash cycle followed by a70% ethanol wash cycle. Filter mats were allowed to air dry and thenplaced into sample bags to which scintillation fluid (Scintiverse II,Fisher Scientific, St. Louis, Mo. or BetaPlate Scintillation Fluid,Pharmacia LKB, Gaithersburg, Md.) was added. Beta emissions of samplesfrom individual tissue culture wells were counted in a LKB Betaplatemodel 1205 scintillation counter (Pharmacia LKB, Gaithersburg, Md.) anddata was expressed as counts per minute of ³H-thymidine incorporatedinto cells from each tissue culture well. Activity of each humaninterleukin-3 preparation or human interleukin-3 variant preparation wasquantitated by measuring cell proliferation (³H-thymidine incorporation)induced by graded concentrations of interleukin-3 or interleukin-3variant. Typically, concentration ranges from 0.05 pM-10⁵ pM arequantitated in these assays. Activity is determined by measuring thedose of interleukin-3 or interleukin-3 variant which provides 50% ofmaximal proliferation [EC₅₀−0.5×(maximum average counts per minute of³H-thymidine incorporated per well among triplicate cultures of allconcentrations of interleukin-3 tested−background proliferation measuredby ³H-thymidine incorporation observed in triplicate cultures lackinginterleukin-3]. This EC₅₀ value is also equivalent to 1 unit ofbioactivity. Every assay was performed with native interleukin-3 as areference standard so that relative activity levels could be assigned.

[0802] Relative biological activities of IL-3 muteins of the presentinvention are shown in Table 1. The Relative Biological Activity of IL-3mutants is calculated by dividing the EC₅₀ of (1-133) hIL-3 by the EC₅₀of the mutant. The Relative Biological Activity may be the average ofreplicate assays. TABLE 1 BIOLOGICAL ACTIVITY OF IL-3 MUTEINS Relative*Plasmid Biological Polypeptide Code Structure Activity Reference (1-133)hIL-3 1 pMON13298 SEQ ID NO. 82 3 pMON13299 SEQ ID NO. 83 2 pMON13300SEQ ID NO. 84 3 pMON13301 SEQ ID NO. 85 2 pMON13302 SEQ ID NO. 86 1.2pMON13303 SEQ ID NO. 87 0.6 pMON13287 SEQ ID NO. 88 26 pMON13288 SEQ IDNO. 89 24 pMON13289 SEQ ID NO. 90 13 pMON13290 SEQ ID NO. 91 20pMON13292 SEQ ID NO. 92 6 pMON13294 SEQ ID NO. 93 3 pMON13295 SEQ ID NO.94 3 pMON13312 SEQ ID NO. 95 4 pMON13313 SEQ ID NO. 96 8 pMON13285 SEQID NO. 259 32 pMON13286 SEQ ID NO. 260 8 pMON13325 SEQ ID NO. 261 8pMON13326 SEQ ID NO. 262 25 pMON13330 SEQ ID NO. 263 19 pMON13329 SEQ IDNO. 406 10 pMON13364 SEQ ID NO. 117 13 pMON13475 SEQ ID NO. 280 7pMON13366 SEQ ID NO. 281 38 pMON13367 SEQ ID NO. 282 36 pMON13368 SEQ IDNO. 278 1.6 pMON13369 SEQ ID NO. 283 10 pMON13370 SEQ ID NO. 284 6pMON13373 SEQ ID NO. 285 12 pMON13374 SEQ ID NO. 286 6 pMON13375 SEQ IDNO. 287 14 pMON13376 SEQ ID NO. 288 0.4 pMON13377 SEQ ID NO. 289 0.4pMON13379 SEQ ID NO. 291 0.9 pMON13380 SEQ ID NO. 279 0.05 pMON13381 SEQID NO. 293 10 pMON13382 SEQ ID NO. 313 38 pMON13383 SEQ ID NO. 294 0.5pMON13384 SEQ ID NO. 295 0.25 pMON13385 SEQ ID NO. 292 1 pMON13387 SEQID NO. 308 32 pMON13388 SEQ ID NO. 296 23 pMON13389 SEQ ID NO. 297 10pMON13391 SEQ ID NO. 298 30 pMON13392 SEQ ID NO. 299 17 pMON13393 SEQ IDNO. 300 32 pMON13394 SEQ ID NO. 301 20 pMON13395 SEQ ID NO. 302 11pMON13396 SEQ ID NO. 303 20 pMON13397 SEQ ID NO. 304 16 pMON13398 SEQ IDNO. 305 36 pMON13399 SEQ ID NO. 306 18 pMON13404 SEQ ID NO. 307 1.3pMON13417 SEQ ID NO. 310 24 pMON13420 SEQ ID NO. 311 19 pMON13421 SEQ IDNO. 312 0.5 pMON13432 SEQ ID NO. 313 10 pMON13400 SEQ ID NO. 317 0.09pMON13402 SEQ ID NO. 318 20 pMON13403 SEQ ID NO. 321 0.03 pMON13405 SEQID NO. 267 9 pMON13406 SEQ ID NO. 264 5 pMON13407 SEQ ID NO. 266 16pMON13408 SEQ ID NO. 269 7 pMON13409 SEQ ID NO. 270 15 pMON13410 SEQ IDNO. 271 0.4 pMON13411 SEQ ID NO. 322 1.2 pMON13412 SEQ ID NO. 323 0.5pMON13413 SEQ ID NO. 324 0.6 pMON13414 SEQ ID NO. 265 4 pMON13415 SEQ IDNO. 268 4 pMON13418 SEQ ID NO. 326 0.5 pMON13419 SEQ ID NO. 325 0.015pMON13422 SEQ ID NO. 272 0.4 pMON13423 SEQ ID NO. 273 0.4 pMON13424 SEQID NO. 274 3 pMON13425 SEQ ID NO. 275 6 pMON13426 SEQ ID NO. 276 >0.0003pMON13429 SEQ ID NO. 277 >0.0002 pMON13440 SEQ ID NO. 319 9 pMON13451SEQ ID NO. 320 0.1 pMON13459 SEQ ID NO. 328 0.003 pMON13416 SEQ ID NO.309 19.9 pMON13428 SEQ ID NO. 327 0.008 pMON13467 SEQ ID NO. 329 0.16pMON13446 SEQ ID NO. 315 21.5 pMON13390 SEQ ID NO. 316 20

[0803] The following assay is used to measure IL-3 mediatedsulfidoleukotriene release from human mononuclear cells.

[0804] IL-3 Mediated Sulfidoleukotriene Release from Human MononuclearCells

[0805] Heparin-containing human blood was collected and layered onto anequal volume of Ficoll-Paque (Pharmacia #17-0840-02) ready to use medium(density 1.077 g/ml.). The Ficoll was warmed to room temperature priorto use and clear 50 ml polystyrene tubes were utilized. The Ficollgradient was spun at 300×g for 30 minutes at room temperature using aH1000B rotor in a Sorvall RT6000B refrigerated centrifuge. The bandcontaining the mononuclear cells was carefully removed, the volumeadjusted to 50 mls with Dulbecco's phosphate-buffered saline (GibcoLaboratories cat. #310-4040PK), spun at 400×g for 10 minutes at 4° C.and the supernatant was carefully removed. The cell pellet was washedtwice with HA Buffer [20 mM Hepes (Sigma #H-3375), 125 mM NaCl (Fisher#S271-500), 5 mM KCl (Sigma #P-9541), 0.5 mM glucose (Sigma #G-5000),0.025% Human Serum Albumin (Calbiochem #126654) and spun at 300×g, 10min., 4° C. The cells were resuspended in HACM Buffer (HA buffersupplemented with 1 mM CaCl₂ (Fisher #C79-500) and 1 mM MgCl₂ (Fisher#M-33) at a concentration of 1×106 cells/ml and 180 μl were transferredinto each well of 96 well tissue culture plates. The cells were allowedto acclimate at 37° C. for 15 minutes. The cells were primed by adding10 μls of a 20× stock of various concentrations of cytokine to each well(typically 100000, 20000, 4000, 800, 160, 32, 6.4, 1.28, 0 fM IL3). Thecells were incubated for 15 minutes at 37° C. Sulfidoleukotriene releasewas activated by the addition of 10 μls of 20×(1000 nM) fmet-leu-phe(Calbiochem #344252) final concentration 50 nM FMLP and incubated for 10minutes at 37° C. The plates were spun at 350×g at 4° C. for 20 minutes.The supernatants were removed and assayed for sulfidoleukotrienes usingCayman's Leukotriene C4 EIA kit (Cat. #420211) according tomanufacturers' directions. Native (15-125) hIL-3 was run as a standardcontrol in each assay.

[0806] Native hIL-3 possesses considerable inflammatory activity and hasbeen shown to stimulate synthesis of the arachidonic acid metabolitesLTC₄, LTD₄, and LTE₄; histamine synthesis and histamine release. Humanclinical trials with native hIL-3 have documented inflammatory responses(Biesma, et al., BLOOD, 80:1141-1148 (1992) and Postmus, et al., J.CLIN. ONCOL., 10:1131-1140 (1992)). A recent study indicates thatleukotrienes are involved in IL-3 actions in vivo and may contributesignificantly to the biological effects of IL-3 treatment (Denzlinger,C., et al., BLOOD, 81:2466-2470 (1993))

[0807] Some muteins of the present invention may have an improvedtherapeutic profile as compared to native hIL-3 or (15-125) hIL-3. Forexample, some muteins of the present invention may have a similar ormore potent growth factor activity relative to native hIL-3 or (15-125)hIL-3 without having a similar or corresponding increase in thestimulation of leukotriene or histamine. These muteins would be expectedto have a more favorable therapeutic profile since the amount ofpolypeptide which needs to be given to achieve the desired growth factoractivity (e. g. cell proliferation) would have a lesser leukotriene orhistamine stimulating effect. In studies with native hIL-3, thestimulation of inflammatory factors has been an undesirable side effectof the treatment. Reduction or elimination of the stimulation ofmediators of inflammation would provide an advantage over the use ofnative hIL-3.

[0808] The pMON13288 polypeptide has demonstrated a more potent growthfactor activity relative to native hIL-3 in the AML 193 cellproliferation assay (EC₅₀=0.8-3.8 pM for pMON13288 and EC₅₀=30.2 pM fornative hIL-3) without demonstrating a corresponding increase in thestimulation of leukotriene C₄ (LTC₄) production and histamine release,i. e., it stimulated LTC₄ production and histamine release with apotency similar to that of native hIL-3 while having an improved abilityto stimulate cell proliferation compared to native hIL-3. Thus with thepMON13288 polypeptide it would be expected that one would be able toproduce a desired therapeutic response, e. g., cell proliferation, withless stimulation of the undesirable inflammatory mediators.

[0809] Some muteins of the present invention have antigenic profileswhich differ from that of native hIL-3. For example, in a competitionELISA with an affinity purified polyclonal goat anti-hIL-3 antibody,native hIL-3 significantly blocked the binding of labeled hIL-3 topolyclonal anti-hIL-3 antibody whereas the pMON13288 polypeptide failedto block the binding of hIL-3 to anti-hIL-3 antibody.

[0810] Table 2 lists the sequences of some oligonucleotides used inmaking the muteins of the present invention.

[0811] Table 3 lists the amino acid sequence of native (15-125) hIL-3(Peptide #1) and the amino acid sequences of some mutant polypeptides ofthe present invention. The sequences are shown with the amino acidnumbering corresponding to that of native hIL-3 [FIG. 1].

[0812] Table 4 lists the nucleotide sequences of some DNA sequenceswhich encode mutant polypeptides of the present invention. TABLE 2OLIGONUCLEOTIDES Oligo #1 Length: 000040 CATGGCTAAC TGCTCTATAATGATCGATGA AATTATACAT [SEQ ID NO:15] Oligo #2 Length: 000045 CTTTAAGTGATGTATAATTT CATCGATCAT TATAGAGCAG TTAGC [SEQ ID NO:16] Oligo #3 Length:000036 CACTTAAAGA GACCACCTGC ACCTTTGCTG GACCCG [SEQ ID NO:17] Oligo #4Length: 000036 GAGGTTGTTC GGGTCCAGCA AAGGTGCAGG TGGTCT [SEQ ID NO:18]Oligo #5 Length: 000036 CACTTAAAGA GACCACCTAA CCCTTTGCTG GACCCG [SEQ IDNO:19] Oligo #6 Length: 000036 GAGGTTGTTC GGGTCCAGCA AAGGGTTAGG TGGTCT[SEQ ID NO:20] Oligo #7 Length: 000036 CACTTAAAGG TTCCACCTGC ACCTTTGCTGGACAGT [SEQ ID NO:21] Oligo #8 Length: 000036 GAGGTTGTTA CTGTCCAGCAAAGGTGCAGG TGGAAC [SEQ ID NO:22] Oligo #9 Length: 000027 AACAACCTCAATGCTGAAGA CGTTGAT [SEQ ID NO:23] Oligo #10 Length: 000018 ATCAACGTCTTGAGCATT [SEQ ID NO:24] Oligo #11 Length: 000027 AACAACCTCA ATTCTGAAGACATGGAT [SEQ ID NO:25] Oligo #12 Length: 000018 ATCCATGTCT TCAGAATT [SEQID NO:26] Oligo #13 Length: 000022 CATGGGAACC ATATGTCAGG AT [SEQ IDNO:27] Oligo #14 Length: 000018 ATCCTGACAT ATGGTTCC [SEQ ID NO:28] Oligo#15 Length: 000016 TGAACCATAT GTCAGG [SEQ ID NO:29] Oligo #16 Length:000024 AATTCCTGAC ATATGGTTCA TGCA [SEQ ID NO:30] Oligo #17 Length:000020 AATTCGAACC ATATGTCAGA [SEQ ID NO:31] Oligo #18 Length: 000020AGCTTCTGAC ATATGGTTCG [SEQ ID NO:32] Oligo #19 Length: 000022 ATCGAACCATATGTCAGATG CA [SEQ ID NO:33] Oligo #20 Length: 000018 TCTGACATATGGTTCGAT [SEQ ID NO:34] Oligo #21 Length: 000036 ATCCTGATGG AACGAAACCTTCGACTTCCA AACCTG [SEQ ID NO:35] Oligo #22 Length: 000027 AAGTCGAAGGTTTCGTTCCA TCAGGAT [SEQ ID NO:36] Oligo #23 Length: 000036 ATCCTGATGGAACGAAACCT TCGAACTCCA AACCTG [SEQ ID NO:37] Oligo #24 Length: 000027AGTTCGAAGG TTTCGTTCCA TCAGGAT [SEQ ID NO:39] Oligo #25 Length: 000024CTCGGATTCG TAAGGSCTGT CAAG [SEQ ID NO:39] Oligo #26 Length: 000024CCTTACGAAT GCGAGCAGGT TTGG [SEQ ID NO:40] Oligo #27 Length: 000024GAGAGCTTCG TAAGGGCTGT CAAG [SEQ ID NO:41] Oligo #28 Length: 000024CCTTACGAAG CTCTCCAGGT TTGG [SEQ ID ND:42] Oligo #29 Length: 000015CACTTAGAAA ATGCA [SEQ ID NO:43] Oligo #30 Length: 000020 TTTTCTAAGTGCTTGACAGC [SEQ ID NO:44] Oligo #31 Length: 000015 AACTTAGAAA ATGCA [SEQID NO:45] Oligo #32 Length: 000020 TTTTCTAAGT TCTTGACAGC [SEQ ID NO:46]Oligo #33 Length: 000048 GGTGATTGGA TGTCGAGAGG GTGCGGCCGT GGCAGAGGGCAGACATGG [SEQ ID NO:47] Oligo #34 Length: 000048 CTGCCCTCTG CCACGGCCGCACCCTCTCGA CATCCAATCA CCATCAAG [SEQ ID NO:48] Oligo #35 Length: 000048GATGATTGGA TGTCGAGAGG GTGCGGCCGT GGGAGAGGGC AGACATGG [SEQ ID NO:49]Oligo #36 Length: 000048 CTGCCCTCTG CCACGGCCGC ACCCTCTCGA CATCCAATCATCATCAAG [SEQ ID NO:50] Oligo #37 Length: 000018 TACGAGATTA CGAAGAAT[SEQ ID NO:51] Oligo #38 Length: 000018 CGTAATCTCG TACCATGT [SEQ IDNO:52] Oligo #39 Length: 000018 TTGGAGATTA CGAAGAAT [SEQ ID NO:53] Oligo#40 Length: 000018 CGTAATCTCC AACCATGT [SEQ ID NO:54] Oligo #41 Length:000019 TGCCTCAATA CCTGATGCA [SEQ ID NO:55] Oligo #42 Length: 000021TCAGGTATTG AGGCAATTCT T [SEQ ID NO:56] Oligo #43 Length: 000026AATTCTTGCC AGTCACCTGC CTTGAT [SEQ ID NO:57] Oligo #44 Length: 000016GCAGGTGACT GGCAAG [SEQ ID NO:58] Oligo #45 Length: 000032 AATTCCGGGAAAAACTGACG TTCTATCTGG TT [SEQ ID NO:59] Oligo #46 Length: 000037CTCAAGGGAA ACCAGATAGA ACGTCAGTTT TTCCCGG [SEQ ID NO:60] Oligo #47Length: 000032 ACCCTTGAGC ACGCGCAGGA ACAACAGTAA TA [SEQ ID NO:61] Oligo#48 Length: 000027 AGCTTATTAC TGTTGTTCCT GCGCGTG [SEQ ID NO:62] Oligo#49 Length: 000032 ACCCTTGAGC AAGCGCAGGA ACAACAGTAA TA [SEQ ID NO:63]Oligo #50 Length: 000027 AGCTTATTAC TGTTGTTCCT GCGCTTG [SEQ ID NO:64]Oligo #51 Length: 000034 GCCGATACCGCGGCATACTCCCACCATTCAGAGA [SEQ IDNO:155] Oligo #52 Length: 000033 GCCGATAAGATCTAAAACGGGTATGGAGAAACA [SEQID NO:156] Oligo #53 ATAGTCTTCCCCAGATATCTAACGCTTGAG [SEQ ID NO:157]Oligo #54 Length: 24 CAATACCTGATGCGTTTTCTAAGT [SEQ ID NO:158] Oligo #55Length: 33 GGTTTCGTTCCATCAGAATGTCCATGTCTTCAG [SEQ ID NO:159] Oligo #165NCOECRV1.REQ Length: 000040 CATGGCTAAC TGCTCTAACA TGATCGATGA AATTATAACA[SEQ ID NO: 162] Oligo #166 NCOECRV4.REQ Length: 000045 CTTTAAGTGTGTTATAATTT CATCGATCAT GTTAGAGCAG TTAGC [SEQ ID NO:163] Oligo #167NCOECRV2.REQ Length: 000036 CACTTAAAGC AGCCACCTTT GCCTTTGCTG GACTTC [SEQID NO:164] Oligo #168 NCOECRV5.REQ Length: 000036 GAGGTTGTTG AAGTCCAGCAAAGGCAAAGC TGGCTG [SEQ ID NO:165] Oligo #169 2D5M6SUP.:REQ Length:000027 AACAACCTCA ATGACGAAGA CATGTCT [SEQ ID NO:166] Oligo #1702D5M6SLO.REQ Length: 000018 AGACATGTCT TCGTCATT [SEQ ID NO:167] Oligo#15(A) Length: 000016 TGAACCATAT GTCAGG [SEQ ID NO:168] Oligo #16(A)Length: 000024 AATTCCTGAC ATATGGTTCA TGCA [SEQ ID NO:169] Oligo #B119ALA1.REQ Length: 000040 CATGGCAAAC TGCTCTATAG CTATCGATGA AATTATACAT[SEQ ID NO:170] Oligo #B2 19ALA4.REQ Length: 000045 CTTTAAGTGATGTATAATTT CATCGATAGC TATAGAGGAG TTTGC [SEQ ID NO:171] Oligo #B319ILE1.REQ Length: 000040 CATGGCAAAC TGCTCTATAA TCATCGATGA AATTATACAT[SEQ ID NO:172] Oligo #B 19ILE4.REQ Length: 000045 CTTTAAGTGA TGTATAATTTCATCGATGAT TATAGAGCAG TTTGC [SEQ ID NO:173] Oligo #B5 49ASP1.REQ Length:000036 ATCCTGGACG AACGAAAGCT TCGAACTCCA AACCTG [SEQ ID NO:174] Oligo #B649ASP4.REQ Length: 000027 AGTTCGAAGG TTTCGTTCGT CCAGGAT [SEQ ID NO:175]Oligo #B7 491LE1.REQ Length: 000036 ATCCTGATCG AACGAAACCT TCGAACTCCAAACCTG [SEQ ID NO:176] Oligo #B8 491LE4.REQ Length: 000027 AGTTCGAAGGTTTCGTTCGA TCAGGAT [SEQ ID NO:177] Oligo #B9 49LEU1.REQ Length: 000036ATCCTGGTGG AACGAAACCT TCGAACTCCA AACCTG [SEQ ID NO:178] Oligo #B1049LEU4.REQ Length: 000027 AGTTCGAAGG TTTCGTTCCA GCAGGAT [SEQ ID NO:179]Oligo #B11 42S45V3.REQ Length: 000027 AACAACCTCA ATTCTGAAGA CGTTGAT [SEQID NO:180] Oligo #B12 42S45V6.REQ Length: 000018 ATCAACGTCT TCAGAATT[SEQ ID NO:181] Oligo #B13 18I23A5H.REQ Length: 000051 CGCGCCATGGCTAAGTGCTC TATAATGATC GATGAAGCAA TACATCACTTA [SEQ ID NO:182] Oligo #B142341HIN3.REQ Length: 000018 CGCGTCGATA AGCTTATT [SEQ ID NO:183] Oligo#B15 2341NCO.REQ Length: 000018 GGAGATATAT CCATGGCT [SEQ ID NO:184]Oligo #B16 2A5M6S0D.REQ Length: 000042 TCGGTCCATC AGAATAGACA TGTCTTCAGCATTGAGGTTG TT [SEQ ID NO:185] Oligo #B17 2A5V6S0D.REQ Length: 000042TCGGTCCATC AGAATAGAAA CGTCTTCAGC ATTGAGGTTG TT [SEQ ID NO:186] Oligo#B18 2D5M6S0D.REQ Length: 000042 TCGGTCCATC AGAATAGACA TGTCTTCGTCATTGAGGTTG TT [SEQ ID NO:187] Oligo #B19 2D5V6S0D.REQ Length: 000042TCGGTCCATC AGAATAGAAA CGTCTTCGTC ATTGAGGTTG TT [SEQ ID NO:188] Oligo#B20 2S5M6S0D.REQ Length: 000042 TCGGTCCATC AGAATAGACA TGTCTTCAGAATTGAGGTTG TT [SEQ ID NO:189] Oligo #B21 2S5V6S0D.REQ Length: 000042TCGGTCCATC AGAATAGAAA CGTCTTCAGA ATTGAGGTTG TT [SEQ ID NO:190 ] Oligo#B22 100ARG3.REQ Length: 000048 CTGCCCTCTG CCACGGCCGC ACCCTCTCGACATCCAATCA TCATCCGT [SEQ ID NO:191] Oligo #B23 100ARG8.REQ Length:000026 AATTCTTGCC AGTCACCTGC ACGGAT [SEQ ID NO:192] Oligo #B24101MET4.REQ Length: 000016 ATGGGTGACT GGCAAG [SEQ ID NO:193] Oligo #B2510R01M8.REQ Length: 000026 AATTCTTGCC AGTCACCCAT ACGGAT [SEQ ID NO:194]Oligo #B26 23ALA1.REQ Length: 000040 CATGGCTAAC TGCTCTATTA TGATCGATGAAGCAATACAT [SEQ ID NO:195] Oligo #B27 23ALA4.REQ Length: 000045CTTTAAGTGA TGTATTGCTT CATCGATCAT AATAGAGCAG TTAGC [SEQ ID NO:196] Oligo#B28 29V2R4S2.REQ Length: 000036 CACTTAAAGG TACCACCTCG CCCTTCCCTG GACCCG[SEQ ID NO:197] Oligo #B29 29V2R4S5.REQ Length: 000036 GAGGTTGTTCGGGTCCAGGG AAGGGCGAGG TGGTAC [SEQ ID NO:198] Oligo #B30 34SER2.REQLength: 000036 CACTTAAAGA GACCACCTGC ACCTTCCCTG GACCCG [SEQ ID NO:199]Oligo #B31 34SER5.REQ Length: 000036 GAGGTTGTTC GGGTCCAGGG AAGGTGCAGGTGGTCT [SEQ ID NO:200] Oligo #B32 42D45M3.REQ Length: 000027 AACAACCTCAATGACGAAGA CATGGAT [SEQ ID NO:201] Oligo #B33 42D45M6.REQ Length: 000018ATCCATGTCT TCGTCATT [SEQ ID NO:202] Oligo #B34 42D45V3.REQ Length:000027 AACAACCTCA ATGACGAAGA CGTCGAT [SEQ ID NO:203] Oligo #B3542D45V6.REQ Length: 000018 ATCGACGTCT TCGTCATT [SEQ ID NO:204] Oligo#B36 42D5M6S3.REQ Length: 000027 AACAACCTCA ATGACGAAGA CATGTCT [SEQ IDNO:205] Oligo #B37 42D5M6S6.REQ Length: 000018 AGACATGTCT TCGTCATT [SEQID NO:206] Oligo #B38 42D5V6S3.REQ Length: 000027 AACAACCTCA ATGACGAAGACGTCTCT [SEQ ID NO:207] Oligo #B39 42D5V6S6.REQ Length: 000018AGAGACGTCT TCGTCATT [SEQ ID NO:208] Oligo #B40 50ASP1.REQ Length: 000036ATCCTGATGG ACCGAAACCT TCGACTTCCA AACCTG [SEQ ID NO:209] Oligo #B4150ASP4.REQ Length: 000027 AAGTCGAAGG TTTCGGTCCA TCAGGAT [SEQ ID NO:210]Oligo #B42 50D56S1.REQ Length: 000036 ATCCTGATGG ACCGAAACCT TCGACTTAGCAACCTG [SEQ ID NO:211] Oligo #B43 56SER5.REQ Length: 000024 CCTTACGAAGCTCTCCAGGT TGCT [SEQ ID NO:212] Oligo #B44 82TRP2.REQ Length: 000018CGTAATCTCT GGCCATGT [SEQ ID NO:213] Oligo #B45 82TRP6.REQ Length: 000018CCAGAGATTA CGAAGAAT [SEQ ID NO:214] Oligo #B46 9E12Q6W1.REQ Length:000032 AATTCCGGGA AAAACTGCAA TTCTATCTGT GG [SEQ ID NO:215] Oligo #B479E12Q6W3.REQ Length: 000037 CTCAAGGGTC CACACATACA ATTGCAGTTT TTCCCGG[SEQ ID NO:216] Oligo #B48 9E12Q6V1.REQ Length: 000032 AATTCCGGGAAAAACTGCAA TTCTATCTGG TT [SEQ ID NO:217] Oligo #B49 9E12Q6V3.REQ Length:000037 CTCAAGGGTA ACCAGATAGA ATTGCAGTTT TTCCCGG [SEQ ID NO:218] Oligo#B50 S09E16V1.REQ Length: 000023 AATTCCGGGA AAAACTGACG TTC [SEQ IDNO:219] Oligo #B51 S09E16V3.REQ Length: 000028 AACCAGATAG AACGTCAGTTTTTCCCGG [SEQ ID NO:220] Oligo #B52 S116VD31.REQ Length: 000023TATCTGGTTA CCCTTGAGTA ATA [SEQ ID NO:221] Oligo #B53 SECR1D33.REQLength: 000018 AGCTTATTAC TTCAAGGGT [SEQ ID NO:222] Oligo #B54S9E2Q6V1.REQ Length: 000023 AATTCCGGGA AAAACTGCAA TTC [SEQ ID NO:223]Oligo #B55 S9E2Q6V3.REQ Length: 000028 AACCAGATAG AATTGCAGTT TTTCCCGG[SEQ ID NO:224] Oligo #B56 Ent338.Lo Length: 61 CGATCATTAT AGAGCAGTTAGCCTTGTCAT CGTCGTCCTT GTAATCAGTT [SEQ ID NO:225] TCTGGATATG C Oligo #B57Ent338.UP Length: 63 CATGGCATAT CCAGAAACTG ATTACAAGGA CCACGATGACAAGGCTAACT [SEQ ID NO:226] GCTCTATAAT GAT 09L2Q8S1.REQ Length: 000032AATTCCGGCT TAAACTGCAA TTCTATCTGT CT [SEQ ID NO:227] 09L2Q6S3.REQ Length:000037 CTCAAGGGTA GACAGATAGA ATTGCAGTTT AAGCCGG [SEQ ID NO:228]117S2.REQ Length: 000032 TCTCTTGAGC AAGCGCAGGA ACAACAGTAA TA [SEQ IDNO:229] 19I0L3A1.REQ Length: 000040 CATGGCAAAC TGCTCTATAA TACTCGATGAAGCAATACAT [SEQ ID NO:230] 19I0L3A4.REQ Length: 000045 CTTTAAGTGATGTATTGCTT CATCGAGTAT TATAGAGGAG TTTGC [SEQ. ID NO.:231] 20P23A1.REQLength: 000040 CATGGCAAAC TGCTCTATAA TGCCAGATGA AGCAATACAT [SEQ. IDNO.:232] 20P23A4.REQ Length: 000045 CTTTAAGTGA TGTATTGCTT CATCTGGCATTATAGAGCAG TTTGC [SEQ. ID NO.:233] 23L1.REQ Length: 000040 CATGGCaAACTGCTCTATAA TGATCGATGA AactgATACAT [SEQ. ID NO.:234] 23L4.REQ Length:000045 CTTTAAGTGA TGTATcagTT CATCGATCAT TATAGAGCAG TTtGC [SEQ. IDNO.:235] 29I45752.REQ Length: 000036 CACTTAAAGA TACCACCTAA CCCTAGCCTGGACAGT [SEQ. ID NO.:236] 29I4S7S5.REQ Length: 000036 GAGGTTAGCACTGTCCAGGC TAGGGTTAGG TGGTAT [SEQ. ID NO.:237] 38A5V6S3.REQ Length:000027 GCTAACCTCA ATTCCGAAGA CGTCTCT [SEQ. ID NO.:238] 38A5V6S6.REQLength: 000018 AGAGACGTCT TCGGAATT [SEQ. ID NO.:239] 50D51S1.REQ Length:000036 ATCCTGATGG ACTCCAACCT TCGAACTCCA AACCTG [SEQ. ID NO.:240]50D51S4.REQ Length: 000027 AGTTCGAAGG TTGGAGTCCA TCAGGAT [SEQ. IDNO.:241] 5VYWPTT3.REQ Length: 000048 GTTCCCTATT GGACGGCCCC TCCCTCTCGAACACCAATCA CGATCAAG [SEQ. ID NO.:242] 5VYWPTT7.REQ Length: 000048CGTGATTGGT GTTCGAGAGG GAGGGGCCGT CCAATAGGGA ACACATGG [SEQ. ID NO.:243]62P3H5S2.REQ Length: 000024 CTCGCATTCC CACATGCTTC TAAG [SEQ. ID NO.:244]62P63H2.REQ Length: 000024 CTCGCATTCC CACATGCTGT CAAG [SEQ. ID NO.:245]62P63H5.REQ Length: 000024 ATGTGGGAAT GCGAGCAGGT TTGG [SEQ. ID NO.:246]65S67Q6.REQ Length: 000020 TTTTCTAATT GCTTAGAAGC [SEQ. ID NO.:247]67Q3.REQ Length: 000015 CAATTAGAAA ATGCA [SEQ. ID NO.:248] 67Q6.REQLength: 00002] TTTTCTAATT GCTTGACAGC [SEQ. ID NO.:249 76P1.REQ Length:000021 TCAGGTATTG AGCCAATTCT T [SEQ. ID NO.:250] 76P5.REQ Length: 000019TGGCTCAATA CCTGATGCA [SEQ. ID NO.:251] 79S2.REQ Length: 000018TCTAATCTCC AACCATGT [SEQ. ID NO.:252] 79S6.REQ Length: 000018 TTGGAGATTAGAAAGAAT [SEQ. ID NO.:253] 9L2Q67S3.REQ Length: 000037 CTCAAGAGAAGACAGATAGA ATTGCAGTTT AAGCGG [SEQ. ID NO.:254] 9LQS1181.REQ Length:000043 AATTCCGGCT TAAACTGCAA TTCTATCTGT CTACCCTTTA ATA [SEQ. ID NO.:256]9LQS1183.REQ Length: 000043 AGCTTATTAA AGGGTAGACA GATAGAATTG CAGTTTAAGCCGG [SEQ. ID NO.:257] S9L2Q6S1.REQ Length: 000043 AATTCCGGCT TAAACTGCAATTCTATCTGT CTACCCTTTA ATA [SEQ. ID NO.:258]

[0813] TABLE 3 POLYPEPTIDES PEPTIDE #1; pMON5988 (Example 43);(15-125)hIL-3         Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr HisLeu [SEQ ID NO:65 ]         15                  20                  25Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Gln Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Gln Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Len Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 Asn Cys Ser Asn Met Ile AspGlu Ile Ile Thr His Leu [SEQ ID NO:65] Lys Gln Pro Pro Leu pro Leu LeuAsp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn LeuArg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn AlaSer Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala ThrAla Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln GlnPEPTIDE #2; pMON13344 (Example 8); (15-125)hIL-3 (18I, 25H, 29R, 32A,37P, 42A and 45V);         Asn Cys Ser Ile Met Ile Asp Glu Ile Ile HisHis Leu [SEQ ID NO:66]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Gln Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #3; pMON13345(Example 9); (15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S and 45M);        Asn Cys Ser Ile Met Ile Asp Glu Ile Ile Thr His Leu [SEQ IDNO:67]         15                  20                  25 Lys Arg ProPro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheSrg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #4; pMON13346(Example 10); (15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42A and 45M);        Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ IDNO:68]         15                  20                  25 Lys Val ProPro Ala pro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #5; pMON13347(Example 12); (15-125)hIL-3 (51R, 55L, 59L, 62V, 67N and 69E);        Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ IDNO:69]         15                  20                  25 Lys Gln ProPro Leu pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Ala Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #6; pMON13348(Example 13); (15-125)hIL-3 (51R, 55L, 60S, 62V, 67N and 69E);        Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ IDNO:704]         15                  20                  25 Lys Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLou Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #7; pMON13349(Example 14); (15-125)hiL-3 (51R, 55T, 59L, 62V, 67H and 69E);        Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ IDNO:71]         15                  20                  25 Lys Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #8; pMON13350(Example 16); (15-125)hIL-3 (73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,105Q);         Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQID NO:72]         15                  20                  25 Lys Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLee Arg Asn Leu Gln Pro Gys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #9: pMON13355(Example 17); (15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A and105Q);         Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQID NO:73]         15                  20                  25 Lys Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #10; pMON13352(Example 19); (15-125)hIL-3 (109E, 116V, 120Q and 123E)         Asn CysSer Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ ID NO:74]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #11; pMON13354(Example 20); (15-125)hIL-3 (109E, 116V, 117S, 120H and 123E);        Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ IDNO:75]         15                  20                  25 Lys Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Ser        105                 110                 115 Leu Glu His Ala GlnGlu Gln Gln         120                 125 PEPTIDE #12; pMON13360(Example 21); (15-125)hIL-3 (73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,105Q, 109E, 116V, 120Q and 123E);         Asn Cys Ser Asn Met Ile AspGlu Ile Ile Thr His Leu [SEQ ID NO:76]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #13; pMON13361(Example 22); (15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 120Q and 123E);         Asn Cys Ser Asn Met Ile AspGlu Ile Ile Thr His Leu [SEQ ID NO:77]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #14; pMON13362(Example 22); (15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 117S, 120H and 123E);         Asn Cys Ser Asn Met IleAsp Glu Ile Ile Thr His Leu [SEQ ID NO:78]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Ser        105                 110                 115 Leu Glu His Ala GlnGlu Gln Gln         120                 125 PEPTIDE #15; pMON13363(Example 24); (15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R,55L, 60S, 62V, 67N and 69E);         Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:79]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #16; pMON13364(Example 25); (15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R,55T, 59L, 62V, 67H and 69E);         Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:80]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Ala Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #17; pMON13365(Example 26 ); (15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R,55L, 59L, 62V, 67N and 69E);         Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:81]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #18; pMON13298(Example 27); Met-Ala-(15-125)hIL-3 (73G, 76A, 79R, 80Q, 87S, 93S, 98I,101A, 105Q, 109E, 116V, 120Q and 123E); Met Ala Asn Cys Ser Asn Met IleAsp Glu Ile Ile Thr His Leu [SEQ ID NO:82]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #19; pMON13299(Example 28); Met-Ala-(15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S, 98T,101A, 105Q, 109E, 116V, 120Q and 123E123E;) Met Ala Asn Cys Ser Asn MetIle Asp Glu Ile Ile Thr His Leu [SEQ ID NO:83]        15                  20                   25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #20; pMON13300(Example 29); Met-Ala-(15-125)hIL-3 (73G, 76A, 79R, 82V, 87S, 93S. 98T,101A, 105Q, 109E, 116V, 117S, 120H and 123E); Met Ala Asn Cys Ser AsnMet Ile Asp Glu Ile Ile Thr His Leu [SEQ ID NO:84]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Ser        105                  110                  115 Leu Glu His AlaGln Ala Gln Gln         120                 125 PEPTIDE #21; pMON13301(Example 30); Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V,51R, 55L, 60S, 62V, 67N, and 69E); Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO:85]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #22; pMON13302(Example 31); Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M,51R, 55T, 59L, 62V, 67H, and 69E): Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO:86]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #23; pMON13303(Example 32); Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M,51R, 55L, 59L, 62V, 67N and 69E); Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO:87]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleIle Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Ala Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #24; pMON13287(Example 33); Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V,51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,105Q, 109E, 116V, 120Q and 123E); Met Ala Asn Cys Ser Ile Met Ile AspGln Ile Ile His His Leu [SEQ ID NO:88]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #25; pMON13288(Example 34); Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32N, 37P, 42S, 45M,51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,105Q, 109E, 116V, 120Q and Met Ala Asn Cys Ser Ile Met Ile Asp Giu IleIle His His Leu [SEQ ID NO:89]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #26; pMON13289(Example 35); Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M,51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A,105Q, 109E, 116V, 120Q, and 123E); Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO:90]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #27; pMON13290(Example 36); Met-Ala-(15-125)hIL-3 (18I, 25H, 29R, 32A, 37P, 42A, 45V,51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 120Q and 123E); Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO:91]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #28; pMON13292(Example 37); Met-Ala-(15-125)hIL-3 (18I, 25H, 29V, 32A, 37S, 42S, 45M,51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 120Q and 123E); Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO:92]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #29; pMON13294Met-Ala-(15-125)hIL-3; (18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T,59L, 62V, 67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E,116V, 117S, 120H and 123E); Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:93 ]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Ser        105                 110                 115 Leu Glu His Ala GlnGlu Gln Gln         120                 125 PEPTIDE #30; pMON13295(Example 39); Met-Ala-(15-125)hIL-3; (18I, 25H, 29V, 32A, 37S, 42S, 45M,51R, 55L, 59L, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 117S, 120H and 123E); Met Ala Asn Cys Ser Ile Met IleAsp Glu Ile Ile His His Leu [SEQ ID NO:94]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Ser        105                 110                 115 Leu Glu His Ala GlnGlu Gln Gln         120                 125 PEPTIDE #31; pMON13312(Example 40); Met-Ala-(15-125)hIL-3; (18I, 25H, 29R, 32N, 37P, 42S, 45M,51R, 55T, 59L, 62V, 67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 120Q and 123E); Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID ND:95]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #32; pMGN13313(Example 41); Met-Ala-(15-125)hIL-3; (18I, 25H, 29R, 32A, 37P, 42A, 45V,51R, 55L, 60S, 62V, 67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A,105Q, 109E, 116V, 117S, 120G and 123E); Met Ala Asn Cys Ser Ile Met IleAsp Glu Ile Ile His His Leu [SEQ ID NO:96]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Val Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Asp Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Ser        105                 110                 115 Leu Glu His Ala GlnGlu Gln Gln         120                 125 PEPTIDE #A3; pMON13285Met-Ala-(15-125)hIL-3; (42D, 45M, 465, 46S, 50D); Met Ala Asn Cys SerAsn Met Ile Asp Glu Ile Ile Thr His Leu ]SEQ ID NO:259]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #A4; pMON13286Met-Ala-(15-125)hIL-3; (42D, 45M, 46S, Met Ala Asn Cys Ser Asn Met IleAsp Gli Ile Ile Thr His Leu [SEQ ID NO:260]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #A5: pMON13325Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 116W); Met Ala Asn Cys Ser Asn MetIle Asp Glu Ile Ile Thr His Leu [SEQ ID NO:261]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Leu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Ala Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Trp Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #A6; pMON13326Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 50D, 116W); Met Ala Asn Cys SerAsn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ ID NO:262]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Asn Asn Leu Arg Arg Pro Asn        45                  50                 55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Ala Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Trp Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #A7; pMON13330Met-Ala-Il-3; (42D, 45M, 46S, 50D, 95R, 98I, 100R, 116W); Met Ala AsnCys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ ID NO:263]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg Arg Pro Ile Ile Ile Arg Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Trp Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #A8; pMON13329Met-Ala-(15-125)hIL-3 (42D, 45M, 46S, 98I, 100R, 116W); Met Ala Asn CysSer Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ ID NO:406]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile Ile Ile Arg Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Trp Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 PEPTIDE #B1Met-Ala-(15-125)hIL-3 pMON13406 Met Ala Asn Cys Ser Ile Ala Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 264]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B2Met-Ala-(15-125)hIL-3 pMON13414 Met Ala Asn Cys Ser Ile Ile Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 265]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B3Met-Ala-(15-125)hIL-3 pMON13407 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu ]SEQ ID NO.: 266]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B4Met-Ala-(15-125)hIL-3 pMON13405 Met Ala Asn Cys Ser Ile Ala Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 267]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #BMet-Ala-(15-125)hIL-3 pMON13415 Met Ala Asn Cys Ser Ile Ile Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 268]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B6Met-Ala-(15-125)hIL-3 pMON13408 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 269]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Ile Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B7Met-Ala-(15-125)hIL3 pMON13409 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 270]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Leu Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE 190 B8Met-Ala-(15-125)hIL-3 pMON13410 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 271]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Asp Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Gln PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B9Met-Ala-(15-125)hIL-3 pMON13422 Met Ala Asn Cys Ser Ile Ala Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 272]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Ile Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His ProIle Ile Ile Lys Ala Gly        90                  95                 100 Asp Trp Gln Gln PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B10Met-Ala-(15-125)hIL-3 pMON13423 Met Ala Asn Cys Ser Ile Ile Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 273]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Ile Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B11Met-Ala-(15-125)hIL-3 pMON13424 Met Ala Asn Cys Ser Ile Ala Ile Asp GluIle Ile His His Leu [SEQ ID NO: 274]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Leu Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Lru Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B12Met-Ala-(15-125)hIL-3 pMON13425 Met Ala Asn Cys Ser Ile Ile Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 275]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Leu Glu Arg Asn Leu Arg Thr Pro Aso        45                  50                  55 Leu Leo Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B13Met-Ala-(15-125)hIL-3 pMON13426 Met Ala Asn Cys Ser Ile Ala Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 276]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Asp Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B14Met-Ala-(15-125)hIL-3 pMON13429 Met Ala Asn Cys Ser Ile Ile Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 277]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Asp IleLeu Asp Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B15Met-Ala-(15-125)hIL-3 pMON13368 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 278]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B16Met-Ala-(15-125)hIL-3 pMON13380 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 279]        15                  20                  25 Lys Val Pro Pro AlaPro Leu Leu Asp Ser Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B17Met-Ala-(15-125)hIL-3 pMON13475 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 280]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B18Met-Ala-(15-125)hIL-3 pMON13366 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 281]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asn        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Gln PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B19Met-Ala-(15-125)hIL-3 pMON13367 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 282]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B20Met-Ala-(15-125)hIL-3 pMON13369 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 283]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B21Met-Ala-(15-125)hIL-3 pMON13370 Met Asp Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 284]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B22Met-Ala-(15-125)hIL-3 pMON13378 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 285]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B23Met-Ala-(15-125)hIL-3 pMON13374 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 286]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B24Met-Ala-(15-125)hIL-3 pMON13375 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 287]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu     119PEPTIDE #B25 Met-Ala-(15-125)hIL-3 pMON13376 Met Asp Asn Cys Ser Ile MetIle Asp Glu Ala Ile His His Leu [SEQ ID NO.: 288]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Gln Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu     119PEPTIDE #B26 Met-Ala-(15-125)hIL-3 pMON13377 Met Ala Asn Cys Ser Ile MetIle Asp Glu Ala Ile His His Leu [SEQ ID NO.: 289]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Trp Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B27Met-Ala-(15-125)hIL-3 pMON13378 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 290]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                115 Leu Glu     119PEPTIDE #B28 Met-Ala-(15-125)hIL-3 pMON13379 Met Ala Asn Cys Ser Ile MetIle Asp Glu Ile Ile His His Leu [SEQ ID NO.: 291]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B29Met-Ala-(15-125)hIL-3 pMON13385 Met Ala Ann Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 292]        15                  20                  25 Lys Val Pro Pro ArgPro Ser Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  95 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B30Met-Ala-(15-125)hIL-3 pMON13381 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 293]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Aso        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Trp Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B31Met-Ala-(15-125)hIL-3 pMON13383 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID ND.: 294]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B32Met-Ala-(15-125)hIL-3 pMON13384 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 295]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B33Met-Ala-(15-125)hIL-3 pMON13388 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 296]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Asp Arg Asn Leu Arg Leu Ser Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B34Met-Ala-(15-125)hIL-3 pMON13389 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 297]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B35Met-Ala-(15-125)hIL-3 pMON13391 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 298]        15                  20                  25 Lys Arg Pro Pro AlaPro Ser Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B36Met-Ala-(15-125)hIL-3 pMON13392 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 299]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B37Met-Ala-(15-125)hIL-3 pMON13393 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 300]        15                  20                  25 Lys Arg Pro Pro AlaPro Ser Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B38Met-Ala-(15-125)hIL-3 pMON13394 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 301]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B39Met-Ala-(15-125)hIL-3 pMON13395 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 302]        15                  20                  25 Lys Val Pro Pro ArgPro Ser Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B40Met-Ala-(15-125)hIL-3 pMON13396 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 303]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Arg Met Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B41Met-Ala-(15-125)hIL-3 pMON13397 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 304]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Trp Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Arg Met Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B42Met-Ala-(15-125)hIL-3 pMON13398 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 305]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B43Met-Ala-(15-125)hIL-3 pMON13399 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 306]        15                  20                  25 Lys Val Pro Pro ArgPro Ser Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Gys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B44Met-Ala-(15-125)hIL-3 pMON13404 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile Ile His His Leu [SEQ ID NO.: 307]        15                  20                      25 Lys Arg Pro ProAla Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu     119PEPTIDE #B45 Met-Ala-(15-125)hIL-3 pMON13387 Met Ala Asn Cys Ser Ile MetIle Asp Glu Ile Ile His His Leu [SEQ ID NO. :308]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Asp Arg Asn Leu Arg Leu Ser Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B46Met-Ala-(15-125)hIL-3 pMON13416 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 309]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B47Met-Ala-(15-125)hIL-3 pMON13417 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 310]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B48Met-Ala-(15-125)hIL-3 pMON13420 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 311]        15                  20                  25 Lys Arg Pro Pro AlaPro Ser Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Val Ser IleLeu Met Asp Arg Asn Leu Arg Leu Ser Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B49Met-Ala-(15-125)hIL-3 pMON13421 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.:331]        15                  20                  25 Lys Arg Pto Pro AlaPro Ser Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Arg Asn Leu Arg Leu Ser Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln Gln         120                 125 PEPTIDE #B50Met-Ala-(15-125)hiL-3 pMON13432 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 312]        15                  20                  25 Lys Arg Pro Pro AlaPro Ser Leu Asp Pro Asn Asn Leu Asn Asp        30                  35                  40 Glu Asp Met Ser IleLeu Met Asp Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B51Met-Ala-(15-125)hIL-3 pMON13382 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 313]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Gln Phe Tyr Leu Trp Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B52Met-Asp-(15-125)hIL-3 pMON13476 Met Asp Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 314]        15                  20                  25 Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Ala        30                  35                  40 Glu Asp Val Asp IleLeu Met Glu Arg Asn Leu Arg Leu Pro Asn        45                  50                  55 Leu Glu Ser Phe ValArg Ala Val Lys Asn Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu pheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #B53Met-Ala-(15-125)hIL-3 pMON13446 Met Ala Tyr Pro Glu Thr Asp Tyr Lys AspAsp Asp Asp Lys Asp [SEQ ID NO.: 315]−14            −10                 −5                   15 Cys Ser IleMet Ile Asp Glu Ile Ile His His Leu Lys Arg Pro                20                  25                  30 Pro Ala ProLeu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp Val                35                  40                  45 Asp Ile LeuMet Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser                50                  55                  60 Phe Val ArgAla Val Lys Asn Leu Glu Asn Ala Ser Gly Ile Glu                65                  70                  75 Ala Ile LeuArg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala                80                  85                  90 Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln                95                  100                 105 Glu Phe ArgGlu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln                110                 115                 120 Ala Gln GluGln Gln                 125 PEPTIDE #54 Met-Ala-(15-125)hIL-3 pMON13390Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp Asp Lys Asn−14            −10                 −5                   15 Cys Ser IleMet Ile Asp Glu Ile Ile His His Leu Lys Arg Pro                20                  25                  30 Pro Asn ProLeu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp Met                35                  40                  45 Asp Ile LeuMet Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala                50                  55                  60 Phe VAl ArgAla Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu                65                  70                  75 Ala Ile LeuArg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala                80                  85                  90 Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln                95                  100                 105 Glu Phe ArgGlu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln                110                 115                 120 Ala Gln GluGln Gln                 125 PEPTIDE #C-2 Met-Ala-(15-125_hIL-3 pMON13400Met Ala Asn Cys Ser Ile Met Pro Asp Glu Ala Ile His His Leu [SEQ ID NO.:317]         15                  20                  25 Lys Ile Pro ProAsn Pro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleL3u Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-3Met-Ala-(15-125)hIL-3 pMON13402 Met Ala Asn Cys Ser Ile Met Ile Asp GluLeu Ile His His Leu [SEQ ID NO.: 318]        15                  20                  25 Lys Ile Pro Pro AsnPro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-10Met-Ala-(15-125)hIL-3 pMON13440 Met Ala Asn Cys Ser Ile Met Ile Asp GluAla Ile His His Leu [SEQ ID NO.: 319]        15                  20                  25 Lys Ile Pro Pro AsnPro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg Thr Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-11Met-Ala-(15-125)hIL-3 pMON13451 Met Ala Asn Cys Ser Ile Ile Leu Asp GluAla Ile His His Leu [SEQ ID NO.: 320]        15                  20                  25 Lys Ile Pro Pro AsnPro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg Thr Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-4Met-Ala-(15-125)hIL-3 pMON13403 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 321]        15                  20                  25 Lys Arg Pro Pro AsePro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Asp Ser Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ProHis Ala Ser Lys Gln Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-5Met-Ala-(15-125)hIL-3 pMON13411 Met Ala Asn Cys Ser Ile Met Ile Asp GlnIle Ile His His Leu [SEQ ID NO.: 322]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-6Met-Ala-(15-125)hIL-3 pMON13412 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 323]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Pro ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Thr Leu        105                 110                 115         118 PEPTIDE#C-7 Met-Ala-(15-125)hIL-3 pMON13413 Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO.: 324]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Ser        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-8Met-Ala-(15-125)hIL-3 pMON13419 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 325]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Asp Ser Asn Leu Leu Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ProHis Ala Ser Lys Gln Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Ser        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-1Met-Ala-(15-125)hIL-3 pMON13418 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 326]        15                  20                  25 Lys Ala Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe VAlArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleLeu Ser Asn Leu Gln Pro Cys Val Pro Tyr        75                  80                  85 Trp Thr Ala Pro ProSer Arg Thr Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-9Met-Ala-(15-125)hIL-3 pMON13428 Met Ala Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu [SEQ ID NO.: 327]        15                  20                  25 Lys Arg Pro Pro AsnPro Leu Leu Asp Pro Asn Asn Leu Asn Ser        30                  35                  40 Glu Asp Met Asp IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleLeu Ser Asn LEu Gln Pro Cys Val Pro Tyr        75                  80                  85 Trp Thr Ala Pro ProSer Arg Thr Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-12Met-Ala-(15-125)hIl-3 pMON13459 Met Ala Asn Cys Ser Ile Met Ile Asp GluLeu Ile His His Leu [SEQ ID NO.: 328]        15                  20                  25 Lys Ile Pro Pro AsnPro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleLeu Ser Asn Leu Gln Pro Cys Val Pro Tyr        75                  80                  85 Trp Thr Ala Pro ProSer Arg Thr Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-13Met-Ala-(15-125)hIL-3 pMON13467 Met Ala Asn Cys Ser Ile Met Ile Asp GluLeu Ile His His Leu [SEQ ID NO.: 329]        15                  20                  25 Lys Ile Pro Pro AsnPro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Ala IleLeu Arg Asn Leu Gln Pro Cys Leu Pro Ser        75                  80                  85 Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Leu Lys Leu Gln Phe Tyr Leu Ser Ser        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125 PEPTIDE #C-14Met-Ala-(15-125)hIL-3 pMON13492 Met Ala Asn Cys Ser Ile Met Ile Asp GluLeu Ile His His Leu [SEQ ID NO.: 330]        15                  20                  25 Lys Ile Pro Pro AsnPro Ser Leu Asp Ser Ala Asn Leu Asn Ser        30                  35                  40 Glu Asp Val Ser IleLeu Met Glu Arg Asn Leu Arg Thr Pro Asn        45                  50                  55 Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser        60                  65                  70 Gly Ile Glu Pro IleLeu Ser Asn Leu Gln Pro Cys Val Pro Tyr        75                  80                  85 Trp Thr Ala Pro ProSer Arg Thr Pro Ile Thr Ile Lys Ala Gly        90                  95                  100 Asp Trp Gln Glu PheArg Glu Lys Leu Thr Phe Tyr Leu Val Thr        105                 110                 115 Leu Glu Gln Ala GlnGlu Gln Gln         120                 125

[0814] TABLE 4 DNA SEQUENCES pMON13287 Met-Ala-(15-125) IL-3 DNAsequence #1ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO:97]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13290 Met-Ala-(15-125) IL-3 DNA sequence #2ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO:98]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCCACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGACGCAATTCTTCCTAATCTCGTACCATCTCTGCCCTCTCCCACGCCCGCACCCTCTCGACATCCAATCACCATCAAGCCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCACGAACAACAG pMON13313 Met-Ala-(15-125) IL-3 DNA sequence #3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO:99]CTGGACCCGAACAACCTCAATGCTGAAGACCTCGATATCCTGATCGAACGAAACCTTCCACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGCTATTGAGCCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTTCCCTTGAGCACGCGCAGGAACAACAG pMON13288 Met-Ala-(15-125) IL-3 DNA sequence #4ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:100]CTGGACCCGAACAACCTCAATTCTGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTSGCAAGAATTCCGGGAAAAACTGAGGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13312 Met-Ala-(15-125) IL-3 DNA sequence #5ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:101]CTGGACCCGAACAACCTCAATTCTGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13294 Met-Ala-(15-125) IL-3 DNA sequence #6ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACGACCTAACCCTTTG [SEQID NO:102]CTGGACCCGAACAACCTCAATTCTGAAGACATGGATATCCTCATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTCCCAACAATTCCGGGAAAAACTGACGTTCTATCTCGTTTCCCTTGAGCACGCCCAGGAACAACAG pMONM13289 Met-Ala-(15-125) IL-3 DNA sequence #7ATGGCTAACTGCTCTATAATCATCGATCAAATTATACATCACTTAAACCTTCCACCTGCACCTTTG [SEQID NO:103]CTGGACAGTAACAACCTCAATTCCGAACACATGCATATCCTCATGCAACGAAACCTTCCACTTCCAAACCTCCTCGCATTCCTAAGCCCTCTCAACAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTCCCCTCTGCCACCCCCCCACCCTCTCCACATCCAATCATCATCAAGGCACGTGACTGGCAAGAATTCCGGCAAAAACTCACGTTCTATCTGCTTACCCTTCACCAAGCCCAGGAACAACAG pMON13292 Met-Ala-(15-125) IL-3 DNA sequence #8ATGGCTAACTCCTCTATAATCATCCATGAAATTATACATCACTTAAACGTTCCACCTGCACCTTTG [SEQID NO:104]CTGGACAGTAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCCACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCTACCATCTCTCCCCTCTGCCACGCCCGCACCCTCTCCACATCCAATCACCATCAACGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTCGTTACCCTTGACCAAGCCCAGGAACAACAG pMON13295 Met-Ala-(15-125) IL-3 DNA sequence #9ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGGTTCCACCTGCACCTTTG [SEQID NO:105]CTGGACAGTAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTTCCCTTGAGCACGCGCAGGAACAACAG pMON13344 (15-125) IL-3 DNA sequence #10AACTGCTCTATAATGATCGATGAAATTATACATGACTTAAAGAGACCACCTGGACCTTTG [SEQ IDNO:106]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGCACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13345 (15-125) IL-3 DNA sequence #11AACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQ IDNO:107]CTGGACCCGAACAACCTCAATTCTGAAGACATGGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13346 (15-125) IL-3 DNA sequence #12AACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGGTTCCACCTGCACCTTTG [SEQ IDNO:108]CTGGACAGTAACAACCTCAATTCCGAAGACATGGATATCCTGATGCAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCCCAGGCTCAACAG pMON13347 (15-125) IL-3 DNA sequence #13AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCACCCACCGCTGCCGCTG [SEQ IDNO:109]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13348 (15-125) IL-3 DNA sequence #14AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:110]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13349 (15-125) IL-3 DNA sequence #15AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:111]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13350 (15-125) IL-3 DNA sequence #16AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:112]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGACAACGCGCAGGCTCAACAG pMON13355 (15-125) IL-3 DNA sequence #17AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:113]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13352 (15-125) IL-3 DNA sequence #18AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:114]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13354 (15-125) IL-3 DNA sequence #19AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:115]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGGGAAAAACTGACGTTCTATCTGGTTTCCCTTGAGCACGCGCAGGAACAACAG pMON13363 (15-125) IL-3 SECRETED DNA sequence #20AACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQ IDNO:116]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13364 (15-125) IL-3 SECRETED DNA sequence #21AACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQ IDNO:117]CTGGACCCGAACAACCTCAATTCTGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCCTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13365 (15-125) IL-3 SECRETED DNA sequence #22AACTGCTCTATAATGATCGATGAAATTATAGATCACTTAAAGGTTCCACCTGCACCTTTG [SEQ IDNO:118]CTGGACAGTAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13360 (15-125) IL-3 SECRETED DNA sequence #23AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:119]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13361 (15-125) IL-3 SECRETED DNA sequence #24AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:120]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13362 (15-125) IL-3 SECRETED DNA sequence #25AACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQ IDNO:121]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTTCCCTTGAGCACGCGCAGGAACAACAG pMON13301 (15-125) IL-3 INTRACELLULAR DNA sequence #26ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO:122]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGCCTGTCAAGAACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATCAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13302 (15-125) IL-3 INTRACELLULAR DNA sequence #27ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:123]CTGGACCGCGAACAACCTCAATTCTGAAGACATGGATATCCTGATGGAACGAAACCTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTACAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCT CCCCTGGCCACGGC GCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATGAATTCCGTCGTAAACTGACCTTCTATCTGAAAACCTTGGAGAACGCGCAGGCTCAACAG pMON13303 (15-125) IL-3 INTRACELLULAR DNA sequence #28ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGGTTCCACCTGCACCTTTG [SEQID NO:124]CTGGACAGTAACAACCTCAATTCCGAAGACATGGATATCCTCATGGAACGAAACCTTCGACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGCAATTGAGAGCATTCTTAAAAATCTCCTGCCATGTCTGCCCCTGGCCACGGCCGCACCCACGCGACATCCAATCCATATCAACGACCGTGACTGGAATGAATTCCGTCCTAAACTCACCTTCTATCTGAAAACCTTGCAGAACCCCCAGGCTCAACAG pMON13298 (15-125) IL-3 INTRACELLULAR DNA sequence #29ATGGCTAACTGCTCTAACATGATCCATGAAATCATCACCCACCTGAAGCAGCCACCCCTCCCGCTG [SEQID NO:125]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCCTCCAAACCTCGAGGCATTCAACCGTGCTGTCAACTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATCTCTCCCCTCTCCCACCGCCCCACCCTCTCCACATCCAATCATCATCAAGGCAGGTGACTGCCAAGAATTCCGGGAAAAACTGACGTTCTATCTGCTTACCCTTGACCAAGCGCAGGAACAACAG pMON13299 (15-125) IL-3 INTRACELLULAR DNA sequence #30ATGGCTAACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQID NO:126]CTGGACTTCAACAACCTCAATGGTCAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAACGCAGGTGACTGGCAAGAATTCCCGGAAAAACTGACGTTCTATCTGGTTACCCTTGACCAAGCGCACGAACAACAG pMON13300 Met-Ala-(15-125) IL-3 INTRACELLULAR DNA sequence #31ATGGCTAACTGCTCTAACATGATCGATGAAATCATCACCCACCTGAAGCAGCCACCGCTGCCGCTG [SEQID NO:127]CTGGACTTCAACAACCTCAATGGTGAAGACCAAGATATCCTGATGGAAAATAACCTTCGTCGTCCAAACCTCGAGGCATTCAACCGTGCTGTCAAGTCTCTGCAGAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCGTACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTTCCCTTGAGCACGCGCAGGAACAACAG DNA sequence #32ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:160]CTGGACCCGAACAACCTCAATTCTGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAACGGCTCTCAAGCACTTAGAAAACGCATCAGCTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #33ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:161]CTGGACCCGAACAACCTCAATTCTGAAGACATGGACATTTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAACGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCOGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B1 pMON13406 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAGCTATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:332]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAACGGCTGTCAAGCACTTAGAAAATGCATCACGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTCCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGCGAAAAACTGACGTTCTATCTGCTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B2 pMON13414 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAATCATCCATGAAATTATACATCACTTAAACAGACCACCTAACCCTTTC [SEQID NO.:333]CTGGACCCGAACAACCTCAATTCCCAACACATCGATATCCTGATGGAACCAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTCTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTCCCACCGCCCCACCCTCTCGACATCCAATCATCATCAAGGCAGCTGACTGCCAAGAATTCCCCGAAAAACTCACGTTCTATCTCGTTACCCTTGACCAACCCCAGGAACAACAG DNA sequence #B3 pMON13407 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:334]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATGCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B4 pMON13405 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAGCTATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:335]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTCCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B5 pMON13415 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAATGATCCATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:336]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCCCAGGAACAACAG DNA sequence #B6 pMON13408 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:337]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATCGAACGAACCTTCGAACTCCAAACCTGCTCCCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACCCCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGCTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B7 pMON13409 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:338]CTGGACCCGAACAACCTCAATTCCGAACACATCGATATCCTGCTGCAACCAAACCTTCGAACTCCAAACCTCCTCCCATTCCTAAGGCCTCTCAACCACTTAGAAAATGCATCACGTATTCAGCCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTCCCACCGCCCCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAACAATTCCGGCAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAACCCCAGGAACAACAG DNA sequence #B8 pMON13410 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCCATGAAATTATACATCACTTAAACAGACCACCTAACCCTTTG [SEQID NO.:339]CTGGACCCGAACAACCTCAATTCCCAACACATGGATATCCTCCACGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTACAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATCTCTCCCCTCTCCCACGCCCGCACCCTCTCCACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACCTTCTATCTCCTTACCCTTGACCAACCGCAGGAACAACAG DNA sequence #B9 pMON13422 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAGCTATCGATGAAATTATACATCACTTAAAGACACCACCTAACCCTTTG [SEQID NO.:340]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATCCTGATCGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACCTTCTATCTGGTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B10 pMON13423 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAATCATCCATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:341]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATCCTGCTGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAACCGCTCTCAACCACTTACAAAATGCATCAGGTATTCAGGCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTGCCACCGCCGCACCCTCTCCACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B11 pMON13424 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAGCTATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:342]CTGGACCCCAACAACCTCAATTCTCAACACGTTGATATCCTGCTCCAACGAAACCTTCCAACTCCAAACCTGCTCGCATTCGTAACCGCTGTCAACCACTTAGAAAATCCATCAGGTATTGACCCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGCCAAAAACTGACGTTCTATCTCGTTACCCTTGACCAACCCCAGGAACAACAG DNA sequence #B12 pMON13425 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAATCATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:343]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATCCTGCTGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCCTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGAGTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B13 pMON13426 Met-Ala-(15-125)IL-3ATGGCAAACTGCTCTATAGCTATCGATGAAATTATACATGAGTTAAAGAGACCACCTAACCCTTTG [SEQID NO.:344]CTGGACCCGAACAACCTCAATTCTGAAGACGTTGATATCCTGGACGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTCCCACCGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTCGCAAGAATTCCGCGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B14 pMON13429 Met-Ala-(15-125)IL-3ATGGCAAACTCCTCTATAATCATCGATGAAATTATACATCACTTAAACAGACCACCTAACCCTTTG [SEQID NO.:345]CTGGACCCGAACAACCTCAATTCTGAAGACGTTATATCCTGGACGAACGAAACCTTCGAACTCCAAACCTCCTCGCATTCGTAACCGCTGTCAACCACTTAGAAAATGCATCAGCTATTGACGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACCGCCCCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B15 pMONM13368 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATTATGATCCATGAACCAATACATCACTTAAACCTTCCACCTCCACCTTTG [SEQID NO.:346]CTGGACAGTAACAACCTCAATTCCCAAGACATGGATATCCTCATCCAACGAAACCTTCCACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCCCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCCGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCCCAGGAACAACAG DNA sequence #B16 pMONM13380 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGGTTCCACCTGCACCTTTG [SEQID NO.:347]CTGGACAGTAACAACCTCAATTCCGAAGACATCGATATCCTCATGGAACGAAACCTTCGACTTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGCAATTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B17 pMON13475 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:348]CTGGACCCGAACAACCTCAATGACGAAGACGTTTCTATTCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGCCAACAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B18 pMON13366 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:349]CTGGACCCGAACAACCTCAATAACGAAGACGTTTCTATTCTGATGCACCCAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B19 pMON13367 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO. :350]CTGGACCCGAACAACCTCAATGCTGAAGACGTTTCTATTCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATCCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGCAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B20 pMON13369 Met-Ala-(15-425)IL-3 42D, 46S, 50DATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGACACCACCTGCACCTTTG [SEQID NO.:351]CTGGACCCGAACAACCTCAATGACGAAGACGTTTCTATTCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B21 pMON13370 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:352]CTGGACCCGAACAACCTCAATGCTGAAGACATGTCTATTCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTCCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGCGAAAAACTCACCTTCTATCTCCTTACCCTTGACCAAGCCCAGGAACAACAG DNA sequence #B22 pMON13373 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATCAAATTATACATCACTTAAAGACACGAGCTGCACCTTTG [SEQID NO.:353]CTGGACCCGAACAACCTCAATCACCAACACATGTCTATTCTCATCGACCCAAACCTTCGAGTTCCAAACCTGGAGACCTTCGTAAGCCCTGTCAACAACTTAGAAAATCCATCAGCTATTGACGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCCGGAAAAACTGACGTTCTATCTCCTTACCCTTGAGCAACCCCAGGAACAACAG DNA sequence #B23 pMON13374 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGACACCACCTCCACCTTTG [SEQID NO.:354]CTGGACCCGAACAACCTCAATGCTGAAGACGTCCATATCCTCATGGAACGAAACCTTCCACTTCCAAACCTGGACAGCTTCGTAAGCGCTGTCAAGAACTTACAAAATCCATCACCTATTCACCCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTGCCACCCCCCCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B24 pMON13375 Met-Ala-(15-119)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAACACACCACCTCCACCTTTG [SEQID NO. 355]CTGGACCCGAACAACCTCAATGCTGAACACGTCGATATCCTGATGGAACGAAACCTTCCACTTCCAAACCTCCACACCTTCCTAACCCCTCTCAACAACTTACAAAATCCATCACCTATTCACCCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAG DNA sequence#B25 pMON13376 Met-Asp-(15-119)IL-3ATGGATAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO. :356]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGCAATTCTATCTGGTTACCCTTGAG DNA sequence#B26 pMON13377 Met-Ala-(15-119)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO. 357]CTGGACCCGAACAACCTCAATGACGAAGACGTCTCTATTCTGATGCACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAACAATTCCGGGAAAAACTGCAATTCTATCTGGTTACCCTTGAC DNA sequence#B27 pMON13378 Met-Asp-(15-119)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGACACCACCTGCACCTTTG [SEQID NO.:358]CTCGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGCCAGGTGACTGGCAAGAATTCCGGCAAAAACTGACGTTCTATCTGGTTACCCTTGAG DNA sequence#B28 pMON13379 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATGGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:359]CTGGACCCGAACAACCTCAATGCTGAAGACGTTTCTATCCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGCAATTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B29 pMON13385 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGGTACCACCTCGCCCTTCC [SEQID NO.:360]CTGGACCCCAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B30 pMON13381 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:361]CTGGACCCGAACAACCTCAATCCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCTGGCCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B31 pMON13383 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATAGATGACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:362]CTGGACCCGAACAACCTCAATGAGGAAGACGTTTCTATTCTGATGGACCGAAACCTTCGAGTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATGCAATCATCATCAAGGCAGGTGAGTGGCAAGAATTCCGGGAAAAACTGCAATTGTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B32 pMON13384 Net-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCCATGAAATTATACATCACTTAAACACACCACCTGCACCTTTG [SEQID NO.:363]CTGGACCCCAACAACCTCAATGCTCAAGACGTCCATATCCTCATGCAACCAAACCTTCGACTTCCAAACCTGGAGAGCTTCCTAAGGCCTGTCAACAACTTAGAAAATCCATCAGGTATTGACCCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTCCCACGGCCGCACCCTCTCCACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGCAATTCTATCTGGTTACCCTTGACCAAGCCCAGGAACAACAG DNA sequence #B33 pMON13388 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAACACACCACCTCCACCTTTG [SEQID NO.:364]CTGGACCCGAACAACCTCAATGCTGAAGACGTCCATATCCTCATGGACCCAAACCTTCGACTTAGCAACCTGGAGACCTTCCTAACCCCTCTCAAGAACTTAGAAAATGCATCACGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACCCCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGCTGACTGCCAAGAATTCCGGGAAAAACTCACGTTCTATCTCCTTACCCTTCACCAACCGCAGGAACAACAG DNA sequence #B34 pMON13389 Net-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:365]CTGGACCCGAACAACCTCAATGAGGAAGACATGGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B35 pMON13391 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGGACCTTCC [SEQID NO.:366]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B36 pMON13392 Met-Ala-(15-125)IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:367]CTGGACCCGAACAACCTCAATGACGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B37 pMON13393 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCCATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTCC [SEQID NO.:368]CTGGACCCGAACAACCTCAATGACGAAGACATGTCTATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGCCAATTCTTCGTAATCTCCAACCATCTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAACGCACGTGACTGCCAACAATTCCGCGAAAAACTGACCTTCTATCTCGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B38 pMON13394 Met-Ala-(15-125) IL-3ATCCCTAACTGCTCTATAATCATCGATGAAATTATACATCACTTAAAGACACCACCTCCACCTTTG [SEQID NO.:369]CTGGACCCGAACAACCTCAATGACCAAGACATGTCTATCCTGATGGAACGAAACCTTCGACTTCCAAACCTCCACACCTTCCTAACCCCTGTCAACAACTTAGAAAATGCATCACCTATTGAGCCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTCCCACGGCCCCACCCTCTCCACATCCAATCATCATCAAGGCAGGTGACTCGCAAGAATTCCCCGAAAAACTGACCTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B39 pMON13395 Met-Ala-(15-125) IL-3ATGGCTAACTCCTCTATAATCATCGATGAAGCAATACATCACTTAAACGTACCACCTCGCCCTTCC [SEQID NO.:370]CTGGACCCGAACAACCTCAATGACCAAGACGTCTCTATCCTGATGCAACCAAACCTTCGACTTCCAAACCTCCAGAGCTTCGTAACGCCTGTCAACAACTTAGAAAATGCATCAGGTATTGACCCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACCCCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGCCAACAATTCCGGCAAAAACTGACCTTCTATCTGGTTACCCTTCAGCAAGCGCAGGAACAACAG DNA sequence #B40 pMON13396 Met-Ala-(15-125)IL-3ATGGCTAACTCCTCTATAATCATCGATGAAATTATACATCACTTAAACAGACCACCTGCACCTTTG [SEQID NO.:371]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTCATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCCGTATGGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B41 pMON13397 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:372]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGAGTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCTGGCCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCCGTATGGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B42 pMON13398 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:373]CTGGACCCGAACAACCTCAATGACGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B43 pMON13399 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGGTACCACCTCGCCCTTCC [SEQID NO.:374]CTGGACCCGAACAACCTCAATGACGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGCCCGCACCCTCTCGACATCCAATCATCATCAAGCCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B44 pMON13404 Met-Ala-(15-119) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:375]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGCAATTCTATCTGGTTACCCTTGAG DNA sequence#B45 pMON13387 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTGCTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAACCGCAGGAACAACAG DNA sequence #B46 pMON13416 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:377]CTGGACCCGAACAACCTCAATGACGAAGACGTCGATTCTCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B47 pMON13287 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:378]CTGGACCGGAACAACCTCAATGACGAAGACGTCATGTCTCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTGGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B48 pMON13420 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGACCTTCC [SEQID NO.:379]CTGGACCCGAACAACCTCAATGACGAACACGTCTCTATCCTGATGGACCGAAACCTTCACTTAGCAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGCGAAAAACTGACGTTCTATCTGGTTACCCTTGACCAAGCGCAGGAACAACAG DNA sequence #B49 pMON13421 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTCC [SEQID NO.:380]CTCGACCCGAACAACCTCAATGACGAAGACATGTCTATCCTGATGGACCGAAACCTTCCACTTAGCAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B50 pMON13432 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTCC [SEQID NO.:381]CTGGACCCGAACAACCTCAATGACGAAGACATGTCTATCCTGATGGACCGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTCCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGCTTACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B51 pMON13382 Met-Ala-(15-125) IL-3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGACACCACCTGCACCTTTG [SEQID NO.:382]CTGGACCCGAACAACCTCAATCCTGAAGACGTCGATATCCTGATCGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCACGTGACTGGCAACAATTCCGGGAAAAACTGCAATTCTATCTGTGGACCCTTGAGCAAGCGCAGGAACAACAG DNA sequence #B52 pMON13476 Met-Asp-(15-125) IL-3ATGGATAACTGCTCTATTATGATCGATGAAGCAATACATCACTTAAAGAGACCACCTGCACCTTTG [SEQID NO.:383]CTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATGGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGAGGTTGTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13400 Met-Ala-(15-125) IL-3 DNA sequence #C2ATGGCTAACTGCTCTATAATGCCAGATGAAGCAATACATCACTTAAAGATACCACCTAACCCTAGC [SEQID NO:384]CTGGACAGTGCTAACCTCAATTCCGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGAGGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13402 Met-Ala-(15-125) IL-3 DNA sequence #C3ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGATACCACCTAACCCTAGC [SEQID NO:385]CTGGACAGTGCTAACCTCAATTCCGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13440 Met-Ala-(15-125) IL-3 DNA sequence #C10ATGGCTAACTGCTCTATTATGATCGATGAAGCAATACATCACTTAAAGATACCACCTAACCCTAGC [SEQID NO:386]CTGGACAGTGCTAACCTCAATTCCGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGAGGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13451 Met-Ala-(15-125) IL-3 DNA sequence #C11ATGGCTAACTGCTCTATAATACTCGATGAAGCAATACATCACTTAAAGATACCACCTAACCCTAGC [SEQID NO:387]CTGGACAGTGCTAACCTCAATTCCGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13403 Met-Ala-(15-125) IL-3 DNA sequence #C4ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:388]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGACTCCAACCTTCGAACTCCAAACCTGCTCGCATTCCCACATGCTGTCAAGCAATTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13419 Met-Ala-(15-125) IL-3 DNA sequence #C8ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:389]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGACTCCAACCTTCGAACTCCAAACCTGCTCGCATTCCCACATCCTTCTAAGCAATTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGCCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13411 Met-Ala-(15-125) IL-3 DNA sequence #C5ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:390]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGCTTAAACTGCAATTCTATCTGTCTACCCTTGAGCAAGCGCAGGAACAACAG pMON13412 Met-Ala-(15-118) IL-3 DNA sequence #C6ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:391]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGCTTAAACTGCAATTCTATCTGTCTACCCTTTAATA pMON13413Met-Ala-(15-125) IL-3 DNA sequence #C7ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:392]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGCTTAAACTGCAATTCTATCTGTCTTCTCTTGAGCAAGCGCAGGAACAACAG pMON13418 Met-Ala-(15-125) IL-3 DNA sequence #C1ATGGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:393]CTGGACCCGAACAACCTCAATTCCCAACACATGGATATCCTCATCGAACCAAACCTTCGAACTCCAAACCTGCTCCCATTCCTAAGGCCTGTCAAGCACTTAGAAAATGCATCACGTATTGACCCAATTCTTTCTAATCTCCAACCATGTGTTCCCTATTCGACCGCCCCTCCCTCTYGAACACCAATCACGATCAAGCCAGCTGACTCGCAAGAATTCCGGCAAAAACTCACCTTCTATCTCGTTACCCTTGACCAACCCCAGGAACAACAC pMON13428 Met-Ala-(15-125) IL-3 DNA sequence #C9ATGGCTAACTGCTGTATAATGATCGATGAAATTATACATCACTTAAAGAGACCACCTAACCCTTTG [SEQID NO:394]CTGGACCCGAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGCCAATTCTTTCTAATCTCCAACCATGTGTTCCCTATTGGACGGCCCCTCCCTCTCGAACACCAATCACGATCAAGGCAGGTGACTGGCAAGAATTCCGGCTTAAACTGGAATTCTATCTGTCTACCCTTGAGCAAGCGCAGGAACAACAG pMON13459 Met-Ala-(15-125) IL-3 DNA sequence #C12ATGGCTAACTGCTCTATAATGATCCATGAAATTATACATCACTTAAAGATACCACCTAACCCTAGC [SEQID NO:395]CTGGACAGTGCTAACCTCAATTCCGAAGACGTCTCTATCCTGATGGAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGCCAATTCTTTCTAATCTCCAACCATGTGTTCCCTATTGGACGGCCCCTCCCTCTCGAACACCAATCACCATCAAGGCAGGTGACTGGCAAGAATTCCGGCTTAAACTGCAATTCTATCTGTCTACCCTTGAGCAAGCGCAGGAACAACAG pMON13467 Met-Ala-(15-125) IL-3 DNA sequence #C13ATGGCTAACTGCTCTATAATCATCCATGAAATTATACATCACTTAAAGATACCACCTAACCCTACC [SEQID NO:396]CTGGACAGTCCTAACCTCAATTCCCAACACCTCTCTATCCTGATCCAACGAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGCCTTAAACTGCAATTCTATCTCTCTTCTCTTGAGCAAGCGGAGGAACAACAG pMON13492 Met-Ala-(15-125) IL-3 DNA sequence #C14ATCGCTAACTGCTCTATAATGATCGATGAAATTATACATCACTTAAAGATACCACCTAACCCTAGC [SEQID NO:397]CTGGACAGTGCTAACCTCAATTCCGAAGACGTCTCTATCCTGATCGAACGAAACCTTCGAACTCCAAACCTCCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGCTATTGAGCCAATTCTTTCTAATCTCCAACCATGTGTTCCCTATTGGACGGCCCCTCCCTCTCGAACACCAATCACGATCAAGGCAGGTSACTGGCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON13446Met-Ala-Tyr-Pro-Glu-Thr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-Ala (15-125)IL-3 DNA sequence #B53ATGGCATATCCAGAAACTGATTACAAGGACCACGATGACAAGGCTAACTGCTCTATAATGATCGAT [SEQID NO:404]GAAATTATACATCACTTAAAGAGACCACCTGCACCTTTGCTGGACCCGAACAACCTCAATGCTGAAGACGTCGATATCCTGATCGAACGAAACCTTCGACTTCCAAACCTGGAGAGCTTCGTAAGGGCTGTCAAGAACTTACAAAATGCATCAGGTATTCAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGCCAGGTGACTGCCAAGAATTCCGGGAAAAACTGACGTTCTATCTGGTTACCCTTGAGCAAGCGCAGGAACAACAG pMON133990Met-Ala-Tyr-Pro-Glu-Thr-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys-Ala (15-125)IL-3 DNA sequence #B54ATGGCATATCCAGAAACTGATTACAAGGACGACGATGACAAGGCTAACTGCTCTATAATGATCGAT [SEQID NO:405]GAAATTATACATCACTTAAAGAGACCACCTAACCCTTTGCTGGACCCCAACAACCTCAATTCCGAAGACATGGATATCCTGATGGAACCAAACCTTCGAACTCCAAACCTGCTCGCATTCGTAAGGGCTGTCAAGCACTTAGAAAATGCATCAGGTATTGAGGCAATTCTTCGTAATCTCCAACCATGTCTGCCCTCTGCCACGGCCGCACCCTCTCGACATCCAATCATCATCAAGGCAGGTGACTGGCAAGAATTCCGGGAAAAACTGACCTTCTATCTCGTTACCCTTGAGCAAGCGCAGGAACAACAG

[0815] Polypeptides corresponding to [SEQ ID NO. 129] comprising (1-133)hIL-3 containing four or more amino acid substitutions can be made usingthe procedures described above and in the following examples by startingwith the appropriate oligonuctiotides and then constructing the DNAencoding the polypeptide and expressing it in an appropriate host cell.In a similar manner polypeptides which correspond to [SEQ ID NO. 130]and contain four or more amino acid substitutions and wherein from 1 to14 amino acids have been sequentially deleted from the N-terminus, orfrom 1 to 15 amino acids have been deleted from the C-terminus ordeletions of amino acids have been made from both the N-terminus and theC-terminus can also be made by following the procedures described aboveand in the following examples, beginning with the appropriate startingmaterials.

[0816] Further details known to those skilled in the art may be found inT. Maniatis, et al., Molecular Cloning, A Laboratory Manual, Cold SpringHarbor Laboratory (1982) and references cited therein, incorporatedherein by reference; and in J. Sambrook, et al., Molecular Cloning, ALaboratory Manual, 2nd edition, Cold Spring Harbor Laboratory (1989) andreferences cited therein, incorporated herein by reference.

[0817] The following examples will illustrate the invention in greaterdetail although it will be understood that the invention is not limitedto these specific examples.

[0818] Amino acids are shown herein by standard one letter or threeletter abbreviations as follows: Abbreviated Designation Amino Acid AAla Alanine C Cys Cysteine D Asp Aspartic acid E Glu Glutamic acid F PhePhenylalanine G Gly Glycine H His Histidine I Ile Isoleucine K LysLysine L Leu Leucine M Met Methionine N Asn Asparagine P Pro Proline QGln Glutamine R Arg Arginine S Ser Serine T Thr Threonine V Val Valine WTrp Tryptophan Y Tyr Tyrosine

[0819] Various other examples will be apparent to the person skilled inthe art after reading the present disclosure without departing from thespirit and scope of the invention. It is intended that all such otherexamples be included within the scope of the appended claims.

[0820] References

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[0857] Olins, P. O. and S. H. Rangwala, Vector for enhanced translationof foreign genes in Escherichia coli, Methods in Enzymology, 185:115-119 (1990).

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[0866] Smith, M. In vitro mutagenesis. Ann. Rev. Genet., 19:423-462(1985).

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EXAMPLE 1

[0880] Construction of pMON 5846 (FIG. 4) which encodes [Met-(1-133)hIL-3 (Arg¹²⁹)]

[0881] A plasmid containing the gene for the cDNA of hIL-3 cloned intopUC18 on an EcoRI to HindIII fragment was obtained from BritishBiotechnology Limited (Cambridge, England). This plasmid was designatedpPO518. The purified plasmid DNA was cleaved by the restrictionendonucleases NheI and BamHI. Approximately 0.5 micrograms of cleavedplasmid DNA was ligated to 1.0 picomoles of a pair of annealedoligonucleotides with the following sequence:5′-CTAGCGATCTTTTAATAAGCTTG-3′ [SEQ ID NO:1]3′-GCTAGAAAATTATTCGAACCTAG-5′ [SEQ ID NO:2]

[0882] The ligation mixture was used to transform competent JM101 cellsto ampicillin resistance. Colonies were picked, and plasmid DNA waspurified and subjected to restriction enzyme analysis. An isolate wasidentified in which the above oligonucleotide sequence had replaced theportion of the gene that encodes the extreme C terminus. Within the newsequence was a new stop codon, TAA, and a recognition site for theenzyme HindIII. The new plasmid was designated pMON5846.

EXAMPLE 2

[0883] (a) Construction of Expression Vector Plasmid pMON2341

[0884] The plasmid pMON2341 was used to supply the particular repliconand expression elements used for construction of many of the plasmidsused to produce hIL-3 and hIL-3 muteins in E. coli. These expressionelements are described in the materials and methods section. pMON2341 isderived from pMON5515 (Olins et al., 1988) and from pMON2429. pMON2429consists of the phage mp18 (Yanisch-Perron et al., 1985) with a BclIfragment carrying the chloramphenicol acetyl transferase (cat) gene frompBR328 (Covarrubias et al., 1981) inserted into the BamHI site. The catgene in pMON2429 has been altered from that in pBR328 by site directedmutagenesis (Kunkel, 1985). The recognition sites for NcoI and EcoRIwhich occur in the native gene were altered so that these tworestriction enzymes no longer recognize these sites. The changes did notalter the protein specified by the gene. Also, an NcoI site wasintroduced at the N-terminus of the coding sequence so that it overlapsthe codon for initiator methionine.

[0885] The steps involved in construction of pMON2341 are listed below:

[0886] (1) The DNAs of pMON5515 and pMON2429 were treated with NcoI andHindIII. The fragments were ligated and used to transform competent E.coli to ampicillin resistance. From these colonies, some were identifiedthat were chloramphenicol resistant. From one of these colonies, plasmidDNA was isolated in which the rat atriopeptigen gene of pMON5515 hadbeen replaced by the NcoI to HindIII fragment containing the cat genefrom pMON2429. This fragment contains the recognition sites for severalrestriction enzymes in the portion derived from the multilinker regionof mp18. The new plasmid was designated pMON2412.

[0887] (2) pMON2412 was treated with the enzyme ClaI which cleaves atone location in the pBR327 derived portion of the DNA. The protrudingends were rendered blunt by treatment with Klenow in the presence ofnucleotide precursors. This DNA was mixed with an isolated 514 bp RsaIfragment derived from pEMBL8 (Dente et al., 1983). This RsaI fragmentcontains the origin of replication of phage f1. This ligation mixturewas used to transform competent E. coli cells to ampicillin resistance.Among the plasmid DNAs isolated from these cells was pMON5578. Thisplasmid has the structure of pMON2412 with the f1 origin region insertedinto the ClaI site. This is illustrated in the Figures and in Olins andRangwala (1990).

[0888] (3) The DNA of pMON5578 was treated with restriction enzymesHindIII and MstII. The DNA was then treated with Klenow enzyme in thepresence of nucleotide precursors to render the ends blunt. This treatedDNA was ligated and used to transform competent E. coli to ampicillinresistance. From the ampicillin resistant colonies, one plasmid wasrecovered from which the portion between HindIII and MstII was absent.This deletion resulted in the removal of sequences from the plasmidwhich are recognized by a number of restriction endonuclease sites. Thenew plasmid was designated pMON5582.

[0889] (4) The DNA of pMON5582 was treated with SstII and BclII andligated in the presence of annealed oligonucleotides with the sequencesshown below. 5′-GGCAACAATTTCTACAAAACACTTGATACTGTAT [SEQ ID NO:3] GAGCAT-3′-CGCCGTTGTTAAAGATGTTTTGTGAACTATGACA [SEQ ID NO:4] TACTCGTA-ACAGTATAATTGCTTCAACAGAACAGATC-3′ TGTCATATTAACGAAGTTGTCTTGT-5′

[0890] This sequence encodes the essential elements of the recA promoterof E. coli including the transcription start site and the lexA repressorbinding site (the operator) (Sancar et al., 1980). The plasmid recoveredfrom the ligation mixes contained this recA promoter in place of the onein pMON5582 (and in pMON5515). The functionality of the recA promoterwas illustrated by Olins and Rangwala (1990). The new plasmid wasdesignated pMON5594.

[0891] (5) To eliminate the single EcoRI site in pMON5594, the DNA wastreated with EcoRI, then with Klenow in the presence of nucleotideprecursors to render the ends blunt and then the DNA was ligated. Fromthis ligation mix a plasmid was recovered whose DNA was not cleaved withEcoRI. This plasmid was designated pMON5630.

[0892] (6) To alter the single recognition site for PstI, plasmidpMON5630 was subjected to site directed mutagensis (Kunkel, 1985). Theoligonucleotide used in this procedure has the sequence shown below.

[0893] 5′-CCATTGCTGCCGGCATCGTGGTC-3′ [SEQ ID NO:5]

[0894] The result of the procedure was to construct pMON2341 whichdiffers from pMON5630 in that the PstI site in the beta-lactamase genewas altered so that PstI no longer recognizes the site. The singlenucleotide change does not alter the amino acid sequence of thebeta-lactamase protein.

[0895] (b) Construction of pMON5847 (FIG. 5) which encodes [Met-(1-133)hIL-3(Arg¹²⁹)]

[0896] Plasmid pMON2341 was used to supply the replicon, promotor,ribosome binding site, transcription terminator and antibioticresistance marker for the plasmids used to produce hIL-3 in E. coli fromcDNA derived hIL-3 genes.

[0897] Plasmid pMON2341 was treated with restriction enzymes NcoI andHindIII. The restriction fragment containing the replication origin waspurified. The DNA of plasmid pMON5846 was treated with NcoI and HindIII.The restriction fragment containing the hIL-3 gene was gel purified.These purified restriction fragments were mixed and ligated. Theligation mixture was used to transform competent JM101 cells toampicillin resistance. Colonies were picked, and plasmid DNA waspurified and analyzed using restriction enzymes. pMON5847 was identifiedas a plasmid with the replicon of pMON2341 and the hIL-3 gene in placeof the chloramphenicol acetyl transferase gene. JM101 cells harboringthis plasmid were cultured in M9 medium and treated with nalidixic acidas described above. Samples of the culture were examined for proteincontent. It was found that this hIL-3 mutein was produced at about 6% oftotal cell protein as measured on Coomassie stained polyacrylamide gels.

EXAMPLE 3

[0898] Construction of pMON5854 (FIG. 7) which Encodes [Met-(1-133)hIL-3(Arg¹²⁹)]

[0899] To increase the accumulation of hIL-3 in E. coli, the codingsequence of the amino terminal portion of the protein was altered tomore closely reflect the codon bias found in E. coli genes that producehigh levels of proteins (Gouy and Gautier, 1982). To change the codingsequence for the amino terminal portion of the gene, a pair of syntheticoligonucleotides were inserted between the NcoI and HpaI sites withinthe coding sequence. About 0.5 micrograms of DNA of the plasmid pMON5847(Example 2) was treated with NcoI and HpaI. This DNA was mixed with anannealed pair of oligonucleotides with the following sequence:5′-CATGGCTCCAATGACTCAGACTACTTCTCTTAAG [SEQ ID NO:6] ACT-    3′-CGAGGTTACTGAGTCTGATGAAGAGAATTC [SEQ ID NO:7] TGA- TCTTGGGTT-3′AGAACCCAA-5′

[0900] The fragments were ligated. The ligation mixture was used totransform competent JM101 to ampicillin resistance. Colonies were pickedinto broth. From the cultures plasmid DNA was made and examined for thepresence of a DdeI site (CTNAG) which occurs in the synthetic sequencebut not between the NcoI and HpaI sites in the sequence of pMON5847. Thenew recombinant plasmid was designated pMON5854. The nucleotide sequenceof the DNA in the coding sequence of the amino terminal portion of thehIL-3 gene in pMON5854 was determined by DNA sequencing and found to bethe same as that of the synthetic oligonucleotide used in ligation.Cultures of JM101 cells harboring this plasmid were grown and treatedwith nalidixic acid to induce production of the hIL-3 mutant protein.Analysis of the proteins on Coomassie gels showed that the accumulationof hIL-3 mutein was about 25% of total cell protein in culturesharboring pMON5854, significantly higher than it was in culturesharboring pMON5847.

EXAMPLE 4 Construction of pMON5887 (FIG. 12) which encodes [Met-(1-125)hIL-3]

[0901] The plasmid DNA of pMON5854 (Example 3) was treated with EcoRIand HindIII and the larger fragment gel was purified. About 0.5microgram of this DNA was ligated to 1 picomole of an annealed pair ofoligonucleotides which encode amino acids 107 through 125 of hIL-3. Thesequences of these oligonucleotides are shown below. EcoRI to HindIII5′-AATTCCGTCGTAAACTGACCTTCTATCTGAAAA- [SEQ ID NO:8]3′-GGCAGCATTTGACTGGAAGATAGACTTTT- [SEQ ID NO:9]CCTTGGAGAACGCGCAGGCTCAACAGTAATA-3′ GGAACCTCTTGCGCGTCCGAGTTGTCATTATTCGA-5′

[0902] After ligation, the DNA was used to transform competent JM101cells to ampicillin resistance. Colonies were picked into broth andplasmid DNA was isolated from each culture. Restriction analysis of theplasmid DNA showed the presence of an EcoRI to HindIII fragment smallerthan that of pMON5854. The nucleotide sequence of the portion of thecoding sequence between the EcoRI and HindIII sites was determined toconfirm the accuracy of the replaced sequence. The new plasmid wasdesignated pMON5887 encoding Met-(1-125) hIL-3 which has the followingamino acid sequence: [SEQ ID NO:10] Met Ala Pro Met Thr Gln Thr Thr SerLeu Lys Thr Ser Trp Val Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr HisLeu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln AspGln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys AsnLeu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile HisIle Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu LysThr Leu Glu Asn Ala Gln Ala Gln Gln

EXAMPLE 5

[0903] Construction of pMON5967 which encodes [Met-Ala-(15-125) hIL-3]

[0904] Plasmid DNA of pMON5887 isolated from E. coli GM48 (dam-) wascleaved with NcoI and ClaI and ligated to 1 picomole of an annealed pairof oligonucleotides, encoding amino acids [Met Ala (15-20) hIL-3]. Thesequence of these oligonucleotides is shown below.5′-CATGGCTAACTGCTCTAACATGAT-3′ [SEQ ID NO:11]    3′-CGATTGACGAGATTGTACTAGC-5′ [SEQ ID NO:12]

[0905] The resulting ligation mix was used to transform competent E.coli JM101 cells to ampicillin resistant colonies. Plasmid DNA wasisolated from these cells and the size of the inserted fragment wasdetermined to be smaller than that of pMON5887 by restriction analysisusing NcoI and NsiI. The nucleotide sequence of the region between NcoIand ClaI was determined and found to be that of the syntheticoligonucleotides. The new plasmid was designated pMON5967 and cellscontaining it were induced for protein production. Sonicated cellpellets and supernatants were used for protein purification andbio-assay.

EXAMPLE 6

[0906] Construction of pMON5978 which Encodes [Met-Ala-(15-125) hIL-3]

[0907] Plasmid DNA of pMON5967 isolated from E. coli GM48(dam-) wascleaved with ClaI and NsiI and ligated to 1 picomole of an annealedassembly of six oligonucleotides encoding hIL-3 amino acids 20-70 (FIG.2). This synthetic fragment encodes three unique restriction sites,EcoRV, XhoI and PstI. The sequence of these oligonucleotides is shown inFIG. 2.

[0908] The resulting ligation mix was used to transform competent E.coli JM101 cells to ampicillin resistant colonies. Plasmid DNA wasisolated and screened with XbaI and EcoRV for the presence of the newrestriction site EcoRV. The DNA sequence of the region between ClaI andNsiI was determined and found to be the same as that of the syntheticoligonucleotides. The new plasmid was designated pMON5978, and cellscontaining it were induced for protein production. Sonicated cellpellets and supernatants were used for protein purification andbio-assay.

[0909] Plasmid pMON5978 encodes [Met-Ala-(15-125) hIL-3] which has thefollowing amino acid sequence: [SEQ ID NO:13] Met Ala Asn Cys Ser AsnMet Ile Asp Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu AspPhe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu ArgArg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala SerAla Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr AlaAla Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe ArgArg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

EXAMPLE 7

[0910] Construction of pMON13356

[0911] Plasmid pMON5988 DNA was digested with restriction enzymes NcoIand EcoRV, and the resulting 4190 base pair NcoI,EcoRV fragment containsthe following genetic elements: beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, pAraBAD promoter, g10L ribosome binding site, lamB secretionleader and the bases encoding amino acids 47-125 of (15-125) hIL-3. The4190 base pair NcoI,EcoRV restriction fragment from pMON5988 was ligatedto the following annealed complementary oligonucleotides from Table (2).

[0912] Oligo #13 [SEQ ID NO:27]

[0913] Oligo #14 [SEQ ID NO:28]

[0914] The ligation reaction mixture was used to transform E. coli K-12strain JM101 and transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth and the size of the inserted fragment was determinedby restriction analysis employing restriction enzymes NcoI and HindIIIin double digest. In the resulting plasmid the 99 bases between the NcoIand EcoRV restriction sites in the (15-125) hIL-3 gene are replaced with22 bases from the above mentioned oligonucleotides. This linker alsocontains a NdeI recognition sequence.

EXAMPLE 8

[0915] Construction of pMON13344

[0916] Plasmid pMON13356 DNA was digested with restriction enzymes NcoIand EcoRV, and the resulting 4190 base pair NcoI,EcoRV fragment containsthe following genetic elements: beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, pAraBAD promoter, g10L ribosome binding site, lamB secretionleader and the bases encoding amino acids 47-125 of (15-125) hIL-3. Thesecond DNA fragment was generated by synthetic gene assembly using thefollowing complementary oligonucleotide pairs that have overlappingends:

[0917] Oligo #1 [SEQ ID NO:15]

[0918] Oligo #2 [SEQ ID NO:16]

[0919] Oligo #3 [SEQ ID NO:17]

[0920] Oligo #4 [SEQ ID NO:18]

[0921] Oligo #9 [SEQ ID NO:23]

[0922] Oligo #10 [SEQ ID NO:24]

[0923] The assembled oligonucleotides create NcoI and EcoRV restrictionends and the DNA sequence that encodes amino acids 15-46 of (15-125)hIL-3 with the following amino acid substitutions: 18I, 25H, 29R, 32A,37P, 42A and 45V. The codons encoding amino acids 15-46 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The 4190 base pairNcoI, EcoRV restriction fragment from pMON13356 was ligated with thepairs of annealed oligonucleotides. The ligation reaction was digestedwith NdeI and subsequently used to transform E. coli K-12 strain JM101.Transformant bacteria were selected on ampicillin-containing plates.Plasmid DNA was isolated from a colony grown in LB broth. The DNAsequence was determined to be that of the oligonucleotides. The plasmid,pMON13344, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #2 [SEQ ID NO:66] Asn Cys Ser Ile Met Ile Asp GluIle Ile His His Leu Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn LeuAsn Ala Glu Asp Val Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn LeuGlu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu SerIle Leu Lys Asn Leu Leu Pro Cys Leo Pro Leu Ala Thr Ala Ala Pro Thr ArgHis Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu ThrPhe Tyr Leo Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[0924] DNA sequence #10 [SEQ ID NO:106] codes for the foregoingpMON13344 polypeptide.

EXAMPLE 9

[0925] Construction of pMON13345

[0926] The 4190 base pair NcoI, EcoRV restriction fragment frompMON13356 was ligated with the following pairs of annealed complementaryoligonucleotides:

[0927] Oligo #1 [SEQ ID NO:15]

[0928] Oligo #2 [SEQ ID NO:16]

[0929] Oligo #5 [SEQ ID NO:19]

[0930] Oligo #6 [SEQ ID NO:20]

[0931] Oligo #11 [SEQ ID NO:25]

[0932] Oligo #12 [SEQ ID NO:26]

[0933] The assembled oligonucleotides create NcoI and EcoRV restrictionends and the DNA sequence that encodes amino acids 15-46 of (15-125)hIL-3 with the following amino acid substitutions: 18I, 25H, 29R, 32N,37P, 42S and 45M. The codons encoding amino acids 15-46 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The ligationreaction was digested with NdeI and used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth. The DNA was sequenced to determine that the sequencewas that of the oligonucleotides. The plasmid, pMON13345, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#3 [SEQ ID NO:67] Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His LeuLys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp MetAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn LeuLeu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His IleLys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys ThrLeu Glu Asn Ala Gln Ala Gln Gln

[0934] DNA sequence #11 [SEQ ID NO:107] codes for the foregoingpMON13345 polypeptide.

EXAMPLE 10

[0935] Construction of pMON13346

[0936] The 4190 base pair NcoI,EcoRV restriction fragment from pMON13356was ligated with the following pairs of annealed complementaryoligonucleotides:

[0937] Oligo #1 [SEQ ID NO:15]

[0938] Oligo #2 [SEQ ID NO:16]

[0939] Oligo #7 [SEQ ID NO:21]

[0940] Oligo #8 [SEQ ID NO:22]

[0941] Oligo #11 [SEQ ID NO:25]

[0942] Oligo #12 [SEQ ID NO:26]

[0943] The assembled oligonucleotides create NcoI and EcoRV restrictionends and the DNA sequence that encodes amino acids 15-46 of (15-125)hIL-3 with the following amino acid substitutions: 18I, 25H, 29V, 32A,37S, 42S and 45M. The codons encoding amino acids 15-46 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The ligationreaction was digested with NdeI and used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth and DNA sequenced to determine that the sequence wasthat of the oligonucleotides. The plasmid, pMON13346, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#4 [SEQ ID NO:68] Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His LeuLys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp MetAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn LeuLeu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His IleLys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys ThrLeu Glu Asn Ala Gln Ala Gln Gln

[0944] DNA sequence #12[SEQ ID NO:108] codes for the foregoing pMON13346polypeptide.

EXAMPLE 11

[0945] Construction of pMON13357

[0946] Plasmid pMON5988 DNA was digested with restriction enzymes EcoRVand NsiI, and the resulting 4218 base pair EcoRV,NsiI fragment containsthe following genetic elements: beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, pAraBAD promoter, g10L ribosome binding site, lamB secretionleader and the bases encoding amino acids 15-46 and 72-125 of (15-125)hIL-3. The 4218 base pair EcoRV,NsiI restriction fragment from pMON5988was ligated to the following annealed complementary oligonucleotides:

[0947] Oligo #19 [SEQ ID NO:33]

[0948] Oligo #20 [SEQ ID NO:34]

[0949] The ligation reaction mixture was used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth, and the size of the inserted fragment was determinedby restriction analysis employing restriction enzymes NcoI and HindIIIin double digest. In the resulting plasmid the 71 bases between theEcoRV and NsiI restriction sites in the (15-125) hIL-3 gene are replacedwith 22 bases from the above mentioned oligonucleotides. This linkeralso contains a NdeI recognition sequence.

EXAMPLE 12

[0950] Construction of pMON13347

[0951] The 4218 base pair EcoRV,NsiI restriction fragment from pMON13357was ligated with the following pairs of annealed complementaryoligonucleotides:

[0952] Oligo #21 [SEQ ID NO:35]

[0953] Oligo #22 [SEQ ID NO:36]

[0954] Oligo #25 [SEQ ID NO:39]

[0955] Oligo #26 [SEQ ID NO:40]

[0956] Oligo #31 [SEQ ID NO:45]

[0957] Oligo #32 [SEQ ID NO:46]

[0958] The assembled oligonucleotides create EcoRV and NsiI restrictionends and the DNA sequence that encodes amino acids 47-71 of (15-125)hIL-3 with the following amino acid substitutions: 51R, 55L, 59L, 62V,67N and 69E. The codons encoding amino acids 47-71 of (15-125) hIL-3 arethose found in the hIL-3 cDNA sequence except at those positions whereamino acid substitutions were made. The ligation reaction was digestedwith NdeI and used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated from a colony grown in LB broth. The DNA was sequenced todetermine that the sequence was that of the oligonucleotides. Theplasmid, pMON13347, encodes the (15-125) hIL-3 variant with thefollowing amino acid sequence: Peptide #5 Asn Cys Ser Asn Met Ile AspGlu Ile [SEQ ID NO:69] Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu LeuAsp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Arg Asn LeuArg Leu Pro Asn Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn AlaSer Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala ThrAla Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[0959] DNA sequence #13 [SEQ ID NO:109] codes for the foregoingpMON13347 polypeptide.

EXAMPLE 13

[0960] Construction of pMON13348

[0961] The 4218 base pair EcoRV,NsiI restriction fragment from pMON13357was ligated with the following pairs of annealed complementaryoligonucleotides:

[0962] Oligo #21 [SEQ ID NO:35]

[0963] Oligo #22 [SEQ ID NO:36]

[0964] Oligo #27 [SEQ ID NO:41]

[0965] Oligo #28 [SEQ ID NO:42]

[0966] Oligo #31 [SEQ ID NO:45]

[0967] Oligo #32 [SEQ ID NO:46]

[0968] The assembled oligonucleotides create EcoRV and NsiI restrictionends and the DNA sequence that encodes amino acids 47-71 of (15-125)hIL-3 with the following amino acid substitutions: 51R, 55L, 60S, 62V,67N and 69E. The codons encoding amino acids 47-71 of (15-125) hIL-3 arethose found in the hIL-3 cDNA sequence except at those positions whereamino acid substitutions were made. The ligation reaction was digestedwith NdeI and used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated from a colony grown in LB broth. The DNA was sequenced todetermine that the sequence was that of the oligonucleotides. Theplasmid, pMON13348, encodes the (15-125) hIL-3 variant with thefollowing amino acid sequence: Peptide #6 Asn Cys Ser Asn Met Ile AspGlu Ile [SEQ ID NO:70] Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu LeuAsp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Arg Asn LeuArg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn AlaSer Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala ThrAla Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[0969] DNA sequence #14 [SEQ ID NO:110] encodes the foregoing pMON13348polypeptide.

EXAMPLE 14

[0970] Construction of pMON13349

[0971] The 4218 base pair EcoRV,NsiI restriction fragment from pMON13357was ligated with the following pairs of annealed complementaryoligonucleotides:

[0972] Oligo #23 [SEQ ID NO:37]

[0973] Oligo #24 [SEQ ID NO:38]

[0974] Oligo #25 [SEQ ID NO:39]

[0975] Oligo #26 [SEQ ID NO:40]

[0976] Oligo #29 [SEQ ID NO:43]

[0977] Oligo #30 [SEQ ID NO:44]

[0978] The assembled oligonucleotides create EcoRV and NsiI restrictionends and the DNA sequence that encodes amino acids 47-71 of (15-125)hIL-3 with the following amino acid substitutions: 51R, 55T, 59L, 62V,67H and 69E. The codons encoding amino acids 47-71 of (15-125) hIL-3 arethose found in the hIL-3 cDNA sequence except at those positions whereamino acid substitutions were made. The ligation reaction was digestedwith NdeI and used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated from a colony grown in LB broth and the DNA was sequenced todetermine that the sequence was that of the oligonucleotides. Theplasmid, pMON13349, encodes the (15-125) hIL-3 variant with thefollowing amino acid sequence: Peptide #7 Asn Cys Ser Asn Met Ile AspGlu Ile [SEQ ID NO:71] Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu LeuAsp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Arg Asn LeuArg Thr Pro Asn Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn AlaSer Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala ThrAla Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[0979] DNA sequence #15 [SEQ ID NO:111] encodes the foregoing pMON13349polypeptide.

EXAMPLE 15

[0980] Construction of pMON13358

[0981] Plasmid pMON5988 DNA was digested with restriction enzymes NsiIand EcoRI and the resulting 4178 base pair NsiT,EcoRI fragment containsthe following genetic elements: beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, pAraBAD promoter, g10L ribosome binding site, lamB secretionleader and the bases encoding amino acids 15-71 and 106-125 of (15-125)hIL-3. The 4178 base pair NsiT,EcoRI restriction fragment from pMON5988was ligated to the following annealed complementary oligonucleotides.

[0982] Oligo #15 [SEQ ID NO:29]

[0983] Oligo #16 [SEQ ID NO:30]

[0984] The ligation reaction mixture was used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth, and the size of the inserted fragment was determinedby restriction analysis employing restriction enzymes NcoI and HindIIIin double digest. In the resulting plasmid the 111 bases between theNsiI and EcoRI restriction sites in the (15-125) hIL-3 gene are replacedwith 24 bases from the above mentioned oligonucleotides. This linkeralso contains a NdeI recognition sequence.

EXAMPLE 16

[0985] Construction of pMON13350

[0986] The 4178 base pair NsiI,EcoRI restriction fragment from pMON13358was ligated with the following pairs of annealed complementaryoligonucleotides:

[0987] Oligo #41 [SEQ ID NO:55]

[0988] Oligo #42 [SEQ ID NO:56]

[0989] Oligo #39 [SEQ ID NO:53]

[0990] Oligo #40 [SEQ ID NO:54]

[0991] Oligo #35 [SEQ ID NO:49]

[0992] Oligo #36 [SEQ ID NO:50]

[0993] Oligo #43 [SEQ ID NO:57]

[0994] Oligo #44 [SEQ ID NO:58]

[0995] The assembled oligonucleotides create NsiI and EcoRI restrictionends and the DNA sequence that encodes amino acids 72-105 of (15-125)hIL-3 with the following amino acid substitutions: 73G, 76A, 79R, 82Q,87S, 93S, 98I, 101A and 105Q. The codons encoding amino acids 72-105 of(15-125) hIL-3 are those found in the hIL-3 cDNA sequence except atthose positions where amino acid substitutions were made. The ligationreaction was digested with NdeI and used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth. The DNA was sequenced to determine that the sequencewas that of the oligonucleotides. The plasmid, pMON13350, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#8 Asn Cys Ser Asn Met Ile Asp Glu Ile [SEQ ID NO:72] Ile Thr His LeuLys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp GlnAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn LeuGln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile IleLys Ala Gly Asp Trp Gln Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys ThrLeu Glu Asn Ala Gln Ala Gln Gln

[0996] DNA sequence #16 [SEQ ID NO:112] codes for the foregoingpMON13350 polypeptide.

EXAMPLE 17

[0997] Construction of pMON13355

[0998] The 4178 base pair NsiI,EcoRI restriction fragment from pMON13358was ligated with the following pairs of annealed complementaryoligonucleotides:

[0999] Oligo #41 [SEQ ID NO:55]

[1000] Oligo #42 [SEQ ID NO:56]

[1001] Oligo #37 [SEQ ID NO:51]

[1002] Oligo #38 [SEQ ID NO:52]

[1003] Oligo #33 [SEQ ID NO:47]

[1004] Oligo #34 [SEQ ID NO:48]

[1005] Oligo #43 [SEQ ID NO:57]

[1006] Oligo #44 [SEQ ID NO:58]

[1007] The assembled oligonucleotides create NsiI and EcoRI restrictionends and the DNA sequence that encodes amino acids 72-105 of (15-125)hIL-3 with the following amino acid substitutions: 73G, 76A, 79R, 82V,87S, 93S, 98T, 101A and 105Q. The codons encoding amino acids 72-105 of(15-125) hIL-3 are those found in the hIL-3 cDNA sequence except atthose positions where amino acid substitutions were made. The ligationreaction was digested with NdeI and used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth. The DNA was sequenced to determine that the sequencewas that of the oligonucleotides. The plasmid, pMON13355, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#9 Asn Cys Ser Asn Met Ile Asp Glu Ile [SEQ ID NO:73] Ile Thr His LeuLys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp GlnAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Per Gly Ile Glu Ala Ile Leu Arg Asn LeuVal Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr IleLys Ala Gly Asp Trp Gln Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys ThrLeu Glu Asn Ala Gln Ala Gln Gln

[1008] DNA sequence #17 [SEQ ID NO:113] codes for the foregoingpMON13355 polypeptide.

EXAMPLE 18

[1009] Construction of pMON13359

[1010] Plasmid pMON5988 DNA was digested with restriction enzymes EcoRIand HindIII, and the resulting 4225 base pair EcoRI,HindIII fragmentcontains the following genetic elements: beta-lactamase gene (AMP),pBR327 origin of replication, phage f1 origin of replication as thetranscription terminator, pAraBAD promoter, g10L ribosome binding site,lamB secretion leader and the bases encoding amino acids 15-105 of(15-125) hIL-3. The 4225 base pair EcoRI,HindIII restriction fragmentfrom pMON5988 was ligated to the following annealed complementaryoligonucleotides.

[1011] Oligo #17 [SEQ ID NO:31]

[1012] Oligo #18 [SEQ ID NO:32]

[1013] The ligation reaction was used to transform E. coli K-12 strainJM101. Transformant bacteria were selected on ampicillin-containingplates. Plasmid DNA was isolated from a colony grown in LB broth, andthe size of the inserted fragment was determined by restriction analysisemploying restriction enzymes NcoI and HindIII in double digest. In theresulting plasmid the 64 bases between the EcoRI and HindIII restrictionsites in the (15-125) hIL-3 gene are replaced with 20 bases from theabove mentioned oligonucleotides. This linker also contains an NdeIrecognition sequence.

EXAMPLE 19

[1014] Construction of pMON13352

[1015] The 4225 base pair EcoRI,HindIII restriction fragment frompMON13359 was ligated with the following pairs of annealed complementaryoligonucleotides:

[1016] Oligo #45 [SEQ ID NO:59]

[1017] Oligo #46 [SEQ ID NO:60]

[1018] Oligo #49 [SEQ ID NO:63]

[1019] Oligo #50 [SEQ ID NO:64]

[1020] The assembled oligonucleotides create EcoRI and HindIIIrestriction ends and the DNA sequence that encodes amino acids 106-125of (15-125) hIL-3 with the following amino acid substitutions: 109E,116V, 120Q and 123E. The codons encoding amino acids 106-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The ligationreaction was digested with NdeI and used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth. The DNA was sequenced to determine that the sequencewas that of the oligonucleotides. The plasmid, pMON13352, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#10 Asn Cys Ser Asn Met Ile Asp Glu Ile [SEQ ID NO:74] Ile Thr His LeuLys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp GlnAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn LeuLeu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His IleLys Asp Gly Asp Trp Asn Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val ThrLeu Glu Gln Ala Gln Glu Gln Gln

[1021] DNA sequence #18 [SEQ ID NO:114] codes for the foregoingpMON13352 polypeptide.

EXAMPLE 20

[1022] Construction of pMON13354

[1023] The 4225 base pair EcoRI,HindIII restriction fragment frompMON13359 was ligated with the following pairs of annealed complementaryoligonucleotides:

[1024] Oligo #45 [SEQ ID NO:59]

[1025] Oligo #46 [SEQ ID NO:60]

[1026] Oligo #47 [SEQ ID NO:61]

[1027] Oligo #48 [SEQ ID NO:62]

[1028] The assembled oligonucleotides create EcoRI and HindIIIrestriction ends and the DNA sequence that encodes amino acids 106-125of (15-125) hIL-3 with the following amino acid substitutions: 109E,116V, 117S, 120H and 123E. The codons encoding amino acids 106-125 of(15-125) hIL-3 are those found in the hIL-3 cDNA sequence except atthose positions where amino acid substitutions were made. The ligationreaction was digested with NdeI and used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated from a colonygrown in LB broth, and the DNA was sequenced to determine that thesequence was that of the oligonucleotides. The plasmid, pMON13354,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #11 Asn Cys Ser Asn Met Ile Asp Gln Ile [SEQ ID NO:75]Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu AsnGly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu GlnAla Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg HisPro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Glu Lys Leu Thr PheTyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln

[1029] DNA sequence #19 [SEQ ID NO:115] codes for the foregoingpMON13354 polypeptide.

EXAMPLE 21

[1030] Construction of pMON13360

[1031] Plasmid pMON13352 DNA was digested with restriction enzymes NsiIand EcoRI, resulting in a 4178 base pair NsiI,EcoRI fragment. Thegenetic elements derived from pMON13352 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite, lamB secretion leader and the bases encoding amino acids 15-71 and106-125 of (15-125) hIL-3. Plasmid pMON13350 DNA was digested with NsiIand EcoRI. The resulting 111 base pair NsiI, EcoRI fragment encodesamino acids 72-105 of (15-125) hIL-3. The eluted restriction fragmentswere concentrated and desalted using Centricon 30 concentrators. Therestriction fragments were ligated, and the ligation reaction mixturewas used to transform E. coli K-12 strain JM101. Transformant bacteriawere selected on ampicillin-containing plates. Plasmid DNA was isolatedand analyzed by restriction analysis. Clones containing the correctinsert lost a XmnI site as compared with pMON13352. Positive clones wereidentified by the loss of a 615 base pair XmnI fragment. The DNA wassequenced to confirm the correct insert. The resulting (15-125) hIL-3variant has the following amino acid substitutions: 73G, 76A, 79R, 82Q,87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codonsencoding amino acids 72-125 of (15-125) hIL-3 are those found in thehIL-3 cDNA sequence except at those positions where amino acidsubstitutions were made. The plasmid, pMON13360, encodes the (15-125)hIL-3 variant with the following amino acid sequence: Peptide #12 AsnCys Ser Asn Met Ile Asp Glu Ile [SEQ ID NO:76] Ile Thr His Leu Lys GlnPro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp IleLeu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala ValLys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn Leu Gln ProCys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys AlaGly Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu GluGln Ala Gln Glu Gln Gln

[1032] DNA sequence #23 [SEQ ID NO:119] encodes the foregoing pMON13360polypeptide.

EXAMPLE 22

[1033] Construction of pMON13361

[1034] Plasmid pMON13352 DNA was digested with restriction enzymes NsiIand EcoRI, resulting in a 4178 base pair NsiI,EcoRI fragment. Thegenetic elements derived from pMON13352 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite, lamB secretion leader and the bases encoding amino acids 15-71 and106-125 of (15-125) hIL-3. Plasmid pMON13355 DNA was digested with NsiIand EcoRI. The resulting 111 base pair NsiI, EcoRI fragment encodesamino acids 72-105 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Clones containing the correct insertcontained an additional RsaI site which results in a 1200 base pairsRsaI fragment. The DNA was sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E,116V, 120Q and 123E. The codons encoding amino acids 72-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13361, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #13 Asn Cys Ser Asn Met Ile Asp Glu Ile [SEQ IDNO:77] Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro AsnLeu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys LeuThr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1035] DNA sequence #24 [SEQ ID NO:120] codes for the foregoingpMON13361 polypeptide.

EXAMPLE 23

[1036] Construction of pMON13362

[1037] Plasmid pMON13354 DNA was digested with restriction enzymes NsiIand EcoRI, resulting in a 4178 base pair NsiI,EcoRI fragment. Thegenetic elements derived from pMON13354 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite, lamB secretion leader and the bases encoding amino acids 15-71 and106-125 of (15-125) hIL-3. Plasmid pMON13355 DNA was digested with NsiIand EcoRI. The resulting 111 base pair NsiI, EcoRI fragment encodesamino acids 72-105 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Clones containing the correct insertcontained an additional RsaI site which results in a 1200 base pairsRsaI fragment. The DNA was sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E,116V, 117S, 120H and 123E. The codons encoding amino acids 72-125 of(15-125) hIL-3 are those found in the hIL-3 cDNA sequence except atthose positions where amino acid substitutions were made. The plasmid,pMON13362, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #14 Asn Cys Ser Asn Met Ile Asp Glu Ile [SEQ IDNO:78] Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro AsnLeu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys LeuThr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln

[1038] DNA sequence #25 [SEQ ID NO:121] codes for the foregoingpMON13362 polypeptide.

EXAMPLE 24

[1039] Construction of pMON13363

[1040] Plasmid pMON13344 DNA was digested with restriction enzymes NsiIand EcoRV, resulting in a 4218 base pair NsiI,EcoRV fragment. Thegenetic elements derived from pMON13344 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite, lamB secretion leader and the bases encoding amino acids 15-46 and72-125 of (15-125) hIL-3. Plasmid pMON13348 DNA was digested with NsiIand EcoRV. The resulting 71 base pair NsiI, EcoRV fragment encodes aminoacids 47-71 of (15-125) hIL-3 The restriction fragments were ligatedwith T4 ligase, and the ligation reaction mixture was used to transformE. coli K-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Clones containing the correct insertcontained an additional DdeI site which results in DdeI restrictionfragments of 806 and 167 base pairs compared to 973 base pairs inpMON13344. The DNA was sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V,67N and 69E. The codons encoding amino acids 15-71 of (15-125) hIL-3 arethose found in the hIL-3 cDNA sequence except at those positions whereamino acid substitutions were made. The plasmid, pMON13363, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#15         Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ IDNO:79] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala GluAsp Val Asp Ile Leu Met Glu Arg Asn Len Arg Leu Pro Asn Leu Glu Ser PheVal Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu LysAsn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro IleHis Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr LeuLys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[1041] DNA sequence #20 [SEQ ID NO:116] codes for the foregoingpMON13363 polypeptide.

EXAMPLE 25

[1042] Construction of pMON13364

[1043] Plasmid pMON13345 DNA was digested with restriction enzymes NsiIand EcoRV, resulting in a 4218 base pair NsiI,EcoRV fragment. Thegenetic elements derived from pMON13345 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite, lamb secretion leader and the bases encoding amino acids 15-46 and72-125 of (15-125) hIL-3. Plasmid pMON13349 DNA was digested with NsiIand EcoRV. The resulting 71 base pair NsiI, EcoRV fragment encodes aminoacids 47-71 of (15-125) hIL-3. The restriction fragments were ligated,and the ligation reaction mixture was used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Clones containing the correct insertcontained an additional DdeT site which results in DdeI restrictionfragments of 806 and 167 base pairs compared to 973 base pairs inpMON13344. The DNA was sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,67H and 69E. The codons encoding amino acids 15-71 of (15-125) hIL-3 arethose found in the hIL-3 cDNA sequence except at those positions whereamino acid substitutions were made. The plasmid, pMON13364, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#16         Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ IDNO:80] Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser GluAsp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala PheVal Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu LysAsn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro IleHis Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr LeuLys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[1044] DNA sequence #21 [SEQ ID NO:117] codes for the foregoingpMON13364 polypeptide.

EXAMPLE 26

[1045] Construction of pMON13365

[1046] Plasmid pMON13346 DNA was digested with restriction enzymes NsiIand EcoRV, resulting in a 4218 base pair NsiI,EcoRV fragment. Thegenetic elements derived from pMON13346 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite, lamB secretion leader and the bases encoding amino acids 15-46 and72-125 of (15-125) hIL-3. Plasmid pMON13347 DNA was digested with NsiIand EcoRV. The resulting 71 base pair NsiI, EcoRV fragment encodes aminoacids 47-71 of (15-125) hIL-3. The restriction fragments were ligated,and the ligation reaction mixture was used to transform E. coli K-12strain JM101. Transformant bacteria were selected onampicillin-containing plates. Clones containing the correct insertcontained an additional DdeI site which results in DdeI restrictionfragments of 806 and 167 base pairs compared to 973 base pairs inpMON13344. The DNA was sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V,67N and 69E. The codons encoding amino acids 15-71 of (15-125) hIL-3 arethose found in the hIL-3 cDNA sequence except at those positions whereamino acid substitutions were made. The plasmid, pMON13365, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#17         Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ IDNO:81] Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser GluAsp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala PheVal Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu LysAsn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro IleHis Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr LeuLys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[1047] DNA sequence #22 [SEQ ID NO:118] codes for the foreging pMON13365polypeptide.

EXAMPLE 27

[1048] Construction of pMON13298

[1049] Plasmid pMON5978 DNA was digested with restriction enzymes NsiIand HindIII, resulting in a 3789 base pair NsiI,HindIII fragment. Thegenetic elements derived from pMON5978 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter, g10L ribosome bindingsite, and the bases encoding amino acids 15-71 of (15-125) hIL-3.Plasmid pMON13360 DNA was digested with NsiI and HindIII. The resulting175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of(15-125) hIL-3. The restriction fragments were ligated, and the ligationreaction mixture was used to transform E. coli K-12 strain JM101.Transformant bacteria were selected on ampicillin-containing plates.Plasmid DNA was isolated, analyzed by restriction analysis, andsequenced to confirm the correct insert. The resulting (15-125) hIL-3variant has the following amino acid substitutions: 73G, 76A, 79R, 82Q,87S, 93S, 98I, 101A, 105Q, 109E, 116V, 120Q and 123E. The codonsencoding amino acids 72-125 of (15-125) hIL-3 are those found in thehIL-3 cDNA sequence except at those positions where amino acidsubstitutions were made. The plasmid, pMON13298, encodes the (15-125)hIL-3 variant with the following amino acid sequence: Peptide #18 MetAla Asn Cys Ser Asn Met Ile Asp Glu Ile Ile Thr His Leu [SEQ ID NO:82]Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp GlnAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn LeuGln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile IleLys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val ThrLeu Glu Gln Ala Gln Glu Gln Gln

[1050] DNA sequence #29 [SEQ ID NO:125] codes for the foregoingpMON13298 polypeptide.

EXAMPLE 28

[1051] Construction of pMON13299

[1052] Plasmid pMON5978 DNA was digested with restriction enzymes NsiIand HindIII, resulting in a 3789 base pair NsiI,HindIII fragment. Thegenetic elements derived from pMON5978 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter, g10L ribosome binding siteand the bases encoding amino acids 15-71 of (15-125) hIL-3. PlasmidpMON13361 DNA was digested with NsiI and HindIII, the resulting 175 basepair NsiI, HindIII fragment encodes amino acids 72-125 of (15-125)hIL-3. The restriction fragments were ligated, and the ligation reactionmixture was used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated, analyzed by restriction analysis, and sequenced to confirm thecorrect insert. The resulting (15-125) hIL-3 variant has the followingamino acid substitutions: 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q,109E, 116V, 120Q and 123E. The codons encoding amino acids 72-125 of(15-125) hIL-3 are those found in the hIL-3 cDNA sequence except atthose positions where amino acid substitutions were made. The plasmid,pMON13299, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #19 Met Ala Asn Cys Ser Asn Met Ile Asp Glu IleIle Thr His Leu [SEQ ID NO:83] Lys Gln Pro Pro Leu Pro Leu Leu Asp PheAsn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg ArgPro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser GlyIle Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala AlaPro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg GluLys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1053] DNA sequence #30 [SEQ ID NO:126] codes for the foregoingpMON13299 polypeptide.

EXAMPLE 29

[1054] Construction of pMON13300

[1055] Plasmid pMON5978 DNA was digested with restriction enzymes NsiIand HindIII, resulting in a 3789 base pair NsiI,HindIII fragment. Thegenetic elements derived from pMON5978 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter, g10L ribosome bindingsite, and the bases encoding amino acids 15-71 of (15-125) hIL-3.Plasmid pMON13362 DNA was digested with NsiI and HindIII. The resulting175 base pair NsiI, HindIII fragment encodes amino acids 72-125 of(15-125) hIL-3. The restriction fragments were ligated, and the ligationreaction mixture was used to transform E. coli K-12 strain JM101.Transformant bacteria were selected on ampicillin-containing plates.Plasmid DNA was isolated, analyzed by restriction analysis, andsequenced to confirm the correct insert. The resulting (15-125) hIL-3variant has the following amino acid substitutions: 73G, 76A, 79R, 82V,87S, 93S, 98T, 10A, 105Q, 109E, 116V, 117S, 120H and 123E. The codonsencoding amino acids 72-125 of (15-125) hIL-3 are those found in thehIL-3 cDNA sequence except at those positions where amino acidsubstitutions were made. The plasmid, pMON13300, encodes the (15-125)hIL-3 variant with the following amino acid sequence: Peptide #20 MetAla Asn Cys Ser Asn Met Ile Asp Gln Ile Ile Thr His Leu [SEQ ID NO:84]Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln Asp GlnAsp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn ArgAla Val Lys Ser Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn LeuVal Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr IleLys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val SerLeu Glu His Ala Gln Glu Gln Gln

[1056] DNA sequence #31 [SEQ ID NO:127] codes for the foregoingpMON13300 polypeptide.

EXAMPLE 30

[1057] Construction of pMON13301

[1058] Plasmid pMON5978 DNA was digested with restriction enzymes NcoIand NsiI, resulting in a 3794 base pair NcoI,NsiI fragment. The geneticelements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327origin of replication, phage f1 origin of replication as thetranscription terminator, precA promoter, g10L ribosome binding site andthe bases encoding amino acids 72-125 of (15-125) hIL-3. PlasmidpMON13363 DNA was digested with NcoI and NsiI. The resulting 170 basepair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125) hIL-3.The restriction fragments were ligated, and the ligation reactionmixture was used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated, analyzed by restriction analysis, and sequenced to confirm thecorrect insert. The resulting (15-125) hIL-3 variant has the followingamino acid substitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L,60S, 62V, 67N and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13301, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #21 Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:85] Lys Arg Pro Pro Ala Pro Leu Leu Asp ProAsn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg LeuPro Asn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser AlaIle Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala AlaPro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg ArgLys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[1059] DNA sequence #26 [SEQ ID NO:122] codes for the foregoingpMON13301 polypeptide.

EXAMPLE 31

[1060] Construction of pMON13302

[1061] Plasmid pMON5978 DNA was digested with restriction enzymes NcoIand NsiI, resulting in a 3794 base pair NcoI,NsiI fragment. The geneticelements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327origin of replication, phage f1 origin of replication as thetranscription terminator, precA promoter, g10L ribosome binding site,and the bases encoding amino acids 72-125 of (15-125) hIL-3. PlasmidpMON13364 DNA was digested with NcoI and NsiI. The resulting 170 basepair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125) hIL-3.The restriction fragments were ligated, and the ligation reactionmixture was used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated, analyzed by restriction analysis, and sequenced to confirm thecorrect insert. The resulting (15-125) hIL-3 variant has the followingamino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T,59L, 62V, 67H and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13302, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #22 Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:86] Lys Arg Pro Pro Asn Pro Leu Leu Asp ProAsn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg ThrPro Asn Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser AlaIle Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala AlaPro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg ArgLys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[1062] DNA sequence #27 [SEQ ID NO:123] codes for the foregoingpMON13302 polypeptide.

EXAMPLE 32

[1063] Construction of pMON13303

[1064] Plasmid pMON5978 DNA was digested with restriction enzymes NcoIand NsiI, resulting in a 3794 base pair NcoI,NsiI fragment. The geneticelements derived from pMON5978 are the beta-lactamase gene (AMP), pBR327origin of replication, phage f1 origin of replication as thetranscription terminator, precA promoter, g10L ribosome binding site,and the bases encoding amino acids 72-125 of (15-125) hIL-3. PlasmidpMON13365 DNA was digested with NcoI and NsiI. The resulting 170 basepair NcoI, NsiI fragment encodes amino acids 15-71 of (15-125) hIL-3.The restriction fragments were ligated, and the ligation reactionmixture was used to transform E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated, analyzed by restriction analysis, and sequenced to confirm thecorrect insert. The resulting (15-125) hIL-3 variant has the followingamino acid substitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L,59L, 62V, 67N and 69E. The codons encoding amino acids 15-71 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13303, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #23 Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:87] Lys Val Pro Pro Ala Pro Leu Leu Asp SerAsn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg LeuPro Asn Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser AlaIle Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala AlaPro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg ArgLys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

[1065] DNA sequence #28 [SEQ ID NO:124] codes for the foregoingpMON13303 polypeptide.

EXAMPLE 33

[1066] Construction of pMON13287

[1067] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13363 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13360 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V,67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,120Q and 123E. The codons encoding amino acids 15-125 of (15-125) hIL-3are those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. The plasmid, pMON13287,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #24 Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile IleHis His Leu [SEQ ID NO:88] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro AsnAsn Leu Asn Ala Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu ProAsn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1068] DNA sequence #1 [SEQ ID NO:97] codes for the foregoing pMON13287polypeptide.

EXAMPLE 34

[1069] Construction of pMON13288

[1070] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13364 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoiI, NsiI fragment encodes amino acids 15-71of (15-125) hIL-3. Plasmid pMON13360 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,67H, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,120Q and 123E. The codons encoding amino acids 15-125 of (15-125) hIL-3are those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. The plasmid, pMON13288,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #25 Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile IleHis His Leu [SEQ ID NO:89] Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro AsnAsn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr ProAsn Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln Glu Gln Gln

[1071] DNA sequence #4 [SEQ ID NO:100] codes for the foregoing pMON13288polypeptide.

EXAMPLE 35

[1072] Construction of pMON13289

[1073] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13365 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13360 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V,67N, 69E, 73G, 76A, 79R, 82Q, 87S, 93S, 98I, 101A, 105Q, 109E, 116V,120Q and 123E. The codons encoding amino acids 15-125 of (15-125) hIL-3are those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. The plasmid, pMON13289,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #26 Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile IleHis His Leu [SEQ ID NO:90] Lys Val Pro Pro Ala Pro Leu Leu Asp Ser AsnAsn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu ProAsn Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1074] DNA sequence #7 [SEQ ID NO:103] codes for the foregoing pMON13289polypeptide.

EXAMPLE 36

[1075] Construction of pMON13290

[1076] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13363 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13361 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V,67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,120Q and 123E. The codons encoding amino acids 15-125 of (15-125) hIL-3are those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. The plasmid, pMON13290,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #27 Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile IleHis His Leu [SEQ ID NO:91] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro AsnAsn Leu Asn Ala Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu ProAsn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1077] DNA sequence #2 [SEQ ID NO:98] codes for the foregoing pMON13290polypeptide.

EXAMPLE 37

[1078] Construction of pMON13292

[1079] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13365 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13361 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V,67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,120Q and 123E. The codons encoding amino acids 15-125 of (15-125) hIL-3are those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. The plasmid, pMON13292,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #28 Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile IleHis His Leu [SEQ ID NO:92] Lys Val Pro Pro Ala Pro Leu Leu Asp Ser AsnAsn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu ProAsn Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1080] DNA sequence #8 [SEQ ID NO:104] codes for the foregoing pMON13292polypeptide.

EXAMPLE 38

[1081] Construction of pMON13294

[1082] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13364 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13362 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,117S, 120H and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13294, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #29 Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:93] Lys Arg Pro Pro Asn Pro Leu Leu Asp ProAsn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg ThrPro Asn Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser GlyIle Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala AlaPro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg GluLys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln

[1083] DNA sequence #6 [SEQ ID NO:102] codes for the foregoing pMON13294polypeptide.

EXAMPLE 39

[1084] Construction of pMON13295

[1085] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13365 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13362 DNA was digested with NsiT andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29V, 32A, 37S, 42S, 45M, 51R, 55L, 59L, 62V,67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,117S, 120H and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13295, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #30 Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:94] Lys Val Pro Pro Ala Pro Leu Leu Asp SerAsn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg LeuPro Asn Leu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser GlyIle Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala AlaPro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg GluLys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln

[1086] DNA sequence #9 [SEQ ID NO:105] codes for the foregoing pMON13295polypeptide.

EXAMPLE 40

[1087] Construction of pMON13312

[1088] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13364 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiI fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13361 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M, 51R, 55T, 59L, 62V,67H, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,120Q and 123E. The codons encoding amino acids 15-125 of (15-125) hIL-3are those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. The plasmid, pMON13312,encodes the (15-125) hIL-3 variant with the following amino acidsequence: Peptide #31 Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile IleHis His Leu [SEQ ID NO:95] Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro AsnAsn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr ProAsn Leu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln

[1089] DNA sequence #5 [SEQ ID NO:101] codes for the foregoing pMON13312polypeptide.

EXAMPLE 41

[1090] Construction of pMON13313

[1091] Plasmid pMON2341 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3619 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, precA promoter and g10L ribosome bindingsite. Plasmid pMON13363 DNA was digested with NcoI and NsiI. Theresulting 170 base pair NcoI, NsiT fragment encodes amino acids 15-71 of(15-125) hIL-3. Plasmid pMON13362 DNA was digested with NsiI andHindIII. The resulting 175 base pair NsiI, HindIII fragment encodesamino acids 72-125 of (15-125) hIL-3. The restriction fragments wereligated, and the ligation reaction mixture was used to transform E. coliK-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis, and sequenced to confirm the correct insert. Theresulting (15-125) hIL-3 variant has the following amino acidsubstitutions: 18I, 25H, 29R, 32A, 37P, 42A, 45V, 51R, 55L, 60S, 62V,67N, 69E, 73G, 76A, 79R, 82V, 87S, 93S, 98T, 101A, 105Q, 109E, 116V,117S, 120H and 123E. The codons encoding amino acids 15-125 of (15-125)hIL-3 are those found in the hIL-3 cDNA sequence except at thosepositions where amino acid substitutions were made. The plasmid,pMON13313, encodes the (15-125) hIL-3 variant with the following aminoacid sequence: Peptide #32 Met Ala Asn Cys Ser Ile Met Ile Asp Glu IleIle His His Leu [SEQ ID NO:96] Lys Arg Pro Pro Ala Pro Leu Leu Asp ProAsn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg LeuPro Asn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser GlyIle Glu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala AlaPro Ser Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg GluLys Leu Thr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln

[1092] DNA sequence #3 [SEQ ID NO:99] codes for the foregoing pMON13313polypeptide.

EXAMPLE 42

[1093] Construction of pMON5987

[1094] Plasmid pMON6458 DNA was digested with restriction enzymes NcoIand HindIII, resulting in a 3940 base pair NcoI,HindIII fragment. Thegenetic elements derived from pMON6458 are the beta-lactamase gene(AMP), pBR327 origin of replication, phage f1 origin of replication asthe transcription terminator, pAraBAD promoter, g10L ribosome bindingsite and lamB secretion leader. Plasmid pMON5978 DNA was digested withNcoI and NsiI. The resulting 170 base pair NcoI, NsiI fragment encodesamino acids 15-71 of (15-125) hIL-3. Plasmid pMON5976 DNA was digestedwith NsiI and HindIII. The resulting 175 base pair NsiI,HindIII fragmentencodes amino acids 72-125 of (15-125) hIL-3. The restriction fragmentswere ligated, and the ligation reaction mixture was used to transform E.coli K-12 strain JM101. Transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated and screened forthe restriction sites EcoRV and NheT and DNA sequenced to confirm thecorrect insert.

EXAMPLE 43

[1095] Construction of pMON5988

[1096] The plasmid DNA of pMON5987 was digested with NheI and EcoRI,resulting in a 3903 base pair NheI, EcoRI fragment. The 3903 base pairNheI, EcoRI fragment was ligated to 1.0 picomoles of the followingannealed oligonucleotides:5′-CTAGCCACGGCCGCACCCACGCGACATCCAATCCATATCAAGGACGGTGACTGGAATG-3′ [SEQ IDNO:131] 3′-GGTGCCGGCGTGGGTGCGCTGTAGGTTAGGTATAGTTCCTGCCACTGACCTTACAATT-5′[SEQ ID NO:132]

[1097] The ligation reaction mixture was used to transform E. coli K-12strain JM101 and transformant bacteria were selected onampicillin-containing plates. Plasmid DNA was isolated and sequenced toconfirm positive clones. This plasmid was constructed to change alanine101 to aspartic acid in the hIL-3 gene (15-125). This plasmid wasdesignated pMON5988.

EXAMPLE 44

[1098] Construction of pMON5853 (FIG. 6) which encodes [Met-(15-133)hIL-3(Arg¹²⁹)]

[1099] Plasmid DNA of pMON5847 (Example 2) was treated with NcoI. Therestriction enzyme was inactivated by heat treatment (65° C. for 10minutes). The DNA was then treated with large fragment of DNA polymeraseI (Klenow) in the presence of all four nucleotide precursors. Thisproduces DNA termini with non-overlapping ends. After 5 minutes at 37°C., the polymerase was inactivated by heat treatment at 65° C. for 10minutes. The DNA was then treated with HpaI, an enzyme which producesnon-overlapping termini. The DNA was ethanol precipitated and ligated.The ligation reaction mixture was used to transform competent JM101cells to ampicillin resistance. Colonies were picked and plasmid DNA wasanalyzed by restriction analysis. A plasmid designated pMON5853 wasidentified as one containing a deletion of the amino terminal 14 codonsof the hIL-3 gene. The DNA sequence for the junction of the ribosomebinding site to the (15-133) hIL-3 gene was determined to be thefollowing: [SEQ ID NO:133] [SEQ ID NO:134]5′-AAGGAGATATATCCATGAACTGCTCTAAC-3′                  M  N  C  S  N

[1100] The lower line contains the one letter code for the amino acidsspecified by the coding sequence of the amino terminus of the 15-133hIL-3 gene. These are methionine, asparagine, cysteine, serine andasparagine.

[1101] When cultures of JM101 cells harboring this plasmid were inducedwith nalidixic acid, it was found that hIL-3 (15-133) accumulated atlevels higher than hIL-3 (pMON5847).

[1102] The plasmid, pMON5853, encodes Met-(15-133) hIL-3 (Arg¹²⁹) whichhas the following amino acid sequence: Met Asn Cys Ser Asn Met Ile Asp[SEQ ID NO:135] Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu LeuAsp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn LeuArg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn AlaSer Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala ThrAla Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln GlnThr Thr Leu Arg Leu Ala Ile Phe

EXAMPLE 45

[1103] Construction of pMON5873 which encodes [Met-(1-133) hIL-3]

[1104] The gene obtained from British Biotechnology, Ltd. specifiedarginine at codon position 129. The amino acid specified in the nativehIL-3 cDNA is serine. To produce a protein with the native sequence atthis position, the portion of the coding sequence between the EcoRI siteat codons 106 and 107 and the NheI site at codons 129 and 130 wasreplaced. Plasmid DNA of pMON5854 (Example 3) and pMON5853 (Example 44)were treated with EcoRI and NheI. The larger fragments of each were gelpurified. These were ligated to a pair of an annealed oligonucleotideswith the following sequences: 5′-AATTCCGTCGTAAACTGACCTTCTATCTGAA [SEQ IDNO:136] AACC- 3′-GGCAGCATTTGACTGGAAGATACACTTTTG [SEQ ID NO:137] G-TTGGAGAACGCGCAGGCTCAACAGACCACTCTGTCG- 3′AACCTCTTGCGCGTCCGAGTTGTCTGGTGAGACAGCG ATC-5′

[1105] The ligation reaction mixtures were used to transform competentJM101 cells to ampicillin resistance. Colonies were picked into brothand grown. Plasmid DNA was isolated and screened for the presence of anew StyI recognition site present in the synthetic DNA and not inpMON5854 and pMON5853. The nucleotide sequence of the gene in the regionbetween EcoRI and NheI was determined and found to be that of thesynthetic oligonucleotides. The new plasmids were designated pMON5873encoding [Met-(1-133) hIL-3] and pMON5872 encoding [Met-(15-133) hIL-3].

[1106] The plasmid, pMON5873, encodes Met-(1-133) hIL-3 which has thefollowing amino acid sequence: Met Ala Pro Met Thr Gln Thr Thr [SEQ IDNO:128] Ser Leu Lys Thr Ser Trp Val Asn Cys Ser Asn Met Ile Asp Glu IleIle Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn Asn Leu AsnGly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro Asn Leu GluAla Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro Thr Arg HisPro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys Leu Thr PheTyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln Thr Thr Leu Ser Leu AlaIle Phe

EXAMPLE 46

[1107] Construction of pMON6458

[1108] Plasmid pMON6525 was digested with restriction enzymes HindIIIand SalIT and the resulting 3172 base pair fragment was isolated from a1% agarose gel by interception onto DEAE membrane. The genetic elementsderived from pMON6525 are the beta-lactamase gene (AMP), pBR327 originof replication, and phage f1 origin of replication as the transcriptionterminator. (The genetic elements derived from plasmid pMON6525 areidentical to those in plasmid pMON2341 which could also be used toconstruct pMON6458.) Plasmid pMON6457 was digested with restrictionenzymes HindIII and SalI and the resulting 1117 base pair fragment wasisolated by PAGE and crush and soak elution. The genetic elementsderived from pMON6457 are the pAraBAD promoter, g10L ribosome bindingsite, lamB secretion leader and the (15-125) hIL-3 gene. The restrictionfragments were ligated and the ligation reaction mixture was used totransform E. coli K-12 strain JM101. Transformant bacteria were selectedon ampicillin-containing plates. Plasmid DNA was isolated and the sizeof the inserted fragment was determined by restriction analysisemploying restriction enzymes NcoI and HindIII in double digest. Clonescontaining the hIL-3 gene (encoding amino acids 15-125) contained a 345base pair NcoI, HindIII restriction fragment. This construct wasdesignated pMON6458. This plasmid was constructed to eliminate an EcoRIrestriction site outside the hIL-3 gene coding region in plasmidpMON6457.

EXAMPLE 47

[1109] Construction of pMON5976 which encodes [Met-(15-125)hIL-3(Ala¹⁰¹)]

[1110] The plasmid DNA of pMON5941 isolated from the dam-E. coli strainGM48 was cleaved with ClaI and NsiI and ligated to 1 picomole of anannealed assembly of six oligonucleotides encoding amino acids 20-70 ofhIL-3 (FIG. 2). This synthetic fragment encodes three unique restrictionsites, EcoRV, XhoI and PstI. The sequence of these oligonucleotides isshown in FIG. 2.

[1111] The resulting ligation mix was used to transform competent E.coli JM101 cells to ampicillin resistant colonies. Plasmid DNA wasisolated and the inserted fragment was determined to have both an EcoRVand NheI site. The nucleotide sequence of the region between ClaI andNsiI was determined and found to be that of the syntheticoligonucleotides. At codons 86-87 of a nucleotide sequence coding for(15-125) hIL-3, an NheI site was introduced. The plasmid with thisalteration was designated pMON5941. This plasmid encodes Met-(15-125)hIL-3 which is altered at position 101 by replacement of aspartate byalanine.

[1112] Plasmid pMON5976 encodes Met-(15-125) hIL-3(Ala¹⁰¹) which has thefollowing amino acid sequence: Met Asn Cys Ser Asn Met Ile Asp [SEQ IDNO:138] Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp PheAsn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg ArgPro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser AlaIle Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala AlaPro Thr Arg His Pro Ile His Ile Lys Ala Gly Asp Trp Asn Glu Phe Arg ArgLys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln

EXAMPLE 48

[1113] Construction of pMON5917 which encodes [Met-(15-88) hIL-3]

[1114] The plasmid DNA of pMON5853 was cleaved with NsiI and HindIII andligated to an annealed pair of oligonucleotides encoding (70-88) hIL-3with a new NheI endonuclease restriction site at codons 86-87. Thesequence of these oligonucleotides is shown in Example 18.

[1115] The ligation mixture was used to transform competent E. coliJM101 cells, and ampicillin resistant colonies were picked. Plasmid DNAisolated from individual colonies was screened for the presence of thenew NheI restriction site. The nucleotide sequence of the substitutedportion was determined and found to be that of the syntheticoligonucleotides. The new plasmid was designated pMON5917 encodingMet-(15-88) hIL-3 containing a new NheI site at codons 86-87.

[1116] Plasmid pMON5917 encodes Met-(15-88) hIL-3 which has thefollowing amino acid sequence: Met Asn Cys Ser Asn Met Ile Asp [SEQ IDNO:139] Glu Ile Ile Thr His Leu Lys Gln Pro Pro Leu Pro Leu Leu Asp PheAsn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg ArgPro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser AlaIle Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala

EXAMPLE 49

[1117] Construction of pmon5941 which encodes [Met-(15-125) hIL-3Ala¹⁰¹]

[1118] The plasmid DNA of pMON5917 was cleaved with NheI and HindIII andligated to two annealed pairs of oligonucleotides which encode aminoacids 86-106 and 107-125 of hIL-3. The sequences of theseoligonucleotides is shown below. NheI to EcoRI5′-CTAGCCACGGCCGCACCCACGCGACATCCAA [SEQ ID NO:140] TCCATATCAAGGCTG-3′-GGTGCCGGCGTGGGTGCGCTGTAGGTTAGGT [SEQ ID NO:141] ATAGTTCCGAC-GTGACTGGAATG-3′ CACTGACCTTACTTAA-5′ EcoRI to HindIII5′-AATTCCGTCGTAAACTGACCTTCTATCTGAA [SEQ ID NO:142] AACCTTGGAGAACGCGCA-3′-CGCAGCATTTGACTGGAACATAGACTTTTGG [SEQ ID NO:143] AACCTCTTGCGCGT-GGCTCAACACTAATA-3′ CCGAGTTGTCATTATTCGA-5′

[1119] The ligation mixture was used to transform competent E. coliJM101 cells to ampicillin resistant colonies. Plasmid DNA was isolatedfrom these cells and the size of the inserted fragment was determined tobe larger by restriction analysis with NcoI and HindIII. The Asp to Ala101 change is encoded on the NheI to EcoRI fragment. The nucleotidesequence of the portion of the coding region between the NheI andHindIII sites was determined and found to be that of the syntheticoligonucleotides. The new plasmid was designated pMON5941.

[1120] The plasmid, pMON5941, encodes Met-(15-125) hIL-3(Ala¹⁰¹) andcontains a new NheI restriction site.

EXAMPLE 50

[1121] Construction of pMON6455

[1122] Plasmid pMON5905 was digested with restriction enzymes HindIIIand NcoI resulting in a 3936 base pair fragment. The genetic elementsderived from pMON5905 are the beta-lactamase gene (AMP), pBR327 originof replication, pAraBAD promoter, g10L ribosome binding site, lamBsecretion leader and phage f1 origin of replication as the transcriptionterminator. The following genetic elements; beta-lactamase gene (AMP),pBR327 origin of replication, g10L ribosome binding site and phage f1origin of replication as the transcription terminator, derived fromplasmid pMON5905 are identical to these in plasmid pMON5594 which couldalso be used to construct pMON6455. The AraBAD promoter is identical tothat described in pMON6235. The lamB signal peptide sequence used inpMON6455 is that shown in FIG. 8 fused to hIL-3 (15-125) at the NcoIsite. Plasmid pMON5887 was digested with restriction enzymes HindIII andNcoI, resulting in a 384 base pair NcoI, HindIII fragment. Therestriction fragments were ligated, and the ligation reaction mixturewas used to transform into E. coli K-12 strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated and the size of the inserted fragment was determined byrestriction analysis employing restriction enzymes NcoI and HindIII indouble digest. Positive clones containing the hIL-3 gene (encoding aminoacids 1-125) contained a 384 base pair NcoI, HindIII restrictionfragment. This construct was designated pMON6455.

EXAMPLE 51

[1123] Construction of pMON6456

[1124] Plasmid pMON5905 was digested with restriction enzymes HindIIIand NcoI resulting in a 3936 base pair fragment. The genetic elementsderived from pMON5905 are the beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, pAraBAD promoter, g10L ribosome binding site and the lamBsecretion leader. Plasmid pMON5871 was digested with restriction enzymesHindIII and NcoI, resulting in a 330 base pair NcoI, HindIII fragment.The genetic element derived from pMON5871 encompassed the bases encodingthe (1-107) hIL-3 gene. The restriction fragments were ligated, and theligation reaction mixture was used to transform E. coli K-12 strainJM101. Transformant bacteria were selected on ampicillin-containingplates. Plasmid DNA was isolated and the size of the inserted fragmentwas determined by restriction analysis employing restriction enzymesNcoI and HindIII in double digest. Clones containing the hIL-3 gene(encoding amino acids 1-107) contained a 330 base pair NcoI, HindIIIrestriction fragment. This construct was designated pMON6456.

EXAMPLE 52

[1125] Construction of pMON6457

[1126] Plasmid pMON6455 DNA grown in E. coli strain GM48 (dam-) wasdigested with restriction enzymes NcoI and ClaI, resulting in a 4263base pair NcoI, ClaI fragment. The restriction fragment was ligated to1.0 picomoles of annealed oligonucleotides with the following sequencecoding for Met Ala 14-20 hIL-3: 5′-CATGGCTAACTGCTCTAACATGAT-3′ [SEQ IDNO:151]     3′-CGATTGACGAGATTGTACTAGC-5′[SEQ ID NO:152]

[1127] The resulting DNA was transformed into E. coli K-12 strain JM101and transformant bacteria were selected on ampicillin-containing plates.Plasmid DNA was isolated and the size of the inserted fragment wasdetermined by restriction analysis employing restriction enzymes XbaIand EcoRI in double digest. Positive clones containing the hIL-3 gene(encoding aa 15-125 of hIL-3) contained a 433 base pair XbaI, EcoRIrestriction fragment. This construct was designated pMON6457. Thisplasmid was constructed to delete the first 14 amino acids of hIL-3. Thecoding sequence of the resulting gene begins as follows: 5′ ATG GCT AACTGC . . . 3′ [SEQ ID NO:153] Met Ala Asn Cys . . . [SEQ ID NO:154]         15

[1128] The first two amino acids (Methionine, Alanine) create an NcoIrestriction site and a signal peptidase cleavage site between the lamBsignal peptide and (15-125) hIL-3. Plasmid pMON6457 encodes (15-125)hIL-3 which has the amino acid sequence designated SEQ ID NO:65.

EXAMPLE 53

[1129] Construction of pMON6235

[1130] One of the DNA fragments used to create this plasmid wasgenerated by site-directed mutagenesis employing PCR techniquesdescribed previously using the following oligonucleotides, Oligo #51[SEQ ID NO:155] and Oligo #52 [SEQ ID NO:156], were used as primers inthis procedure. The template for the PCR reaction was E. coli strainW3110 chromosomal DNA, prepared as described in Maniatis (1982). Theoligonucleotide primers were designed to amplify the AraBAD promoter(Greenfield et al., 1978). The resulting DNA product was digested withthe restriction enzymes SacII and BglII. The reaction mixture waspurified as described previously. Plasmid, pMON5594, DNA was digestedwith SacII and BglII, resulting in a 4416 base pair SacII,BglIIrestriction fragment which contains the following genetic elements;beta-lactamase gene (AMP), pBR327 origin of replication, G10L ribosomebinding site, phage f1 origin of replication as the transcriptionterminator and the chloramphenicol acetyl transferase (cat) gene. The4416 base pair SacII,BglII restriction fragment from pMON5594 wasligated to the PCR-generated SacII, BglII DNA fragment. The ligationmixture was used to transform E. coli K-112 strain JM101. Positiveclones contained a 323 base pair SacII,BglII fragment and were DNAsequenced to confirm that the SacII,BglII fragment was the AraBADpromoter. This construct was designated pMON6235.

EXAMPLE 54

[1131] Construction of pMON5647

[1132] Plasmid pMON5585 [prepared as disclosed in EP 0241446incorporated herein by reference in its entirety] DNA was digested withrestriction enzymes NcoI and HindIII resulting in a 3273 base pairNcoI,HindIII fragment. The genetic elements derived from pMON5585 arethe pBR327 origin of replication, precA promoter, g10L ribosome bindingprotein, bovine somatotropin gene (bST), beta-lactamase gene (AMP) andT7 transcription terminator. Plasmid pMON3267 [prepared as disclosed inEP 0241446 incorporated herein by reference in its entirety] DNA wasdigested with NcoI and HindIII enzymes resulting in a 580 base pairNcoI,HindIII fragment which contains the porcine somatotropin (pST)gene. The restriction fragments were ligated and the ligation reactionmixture was used to transform E. coli strain JM101. Transformantbacteria were selected on ampicillin-containing plates. Plasmid DNA wasisolated, analyzed by restriction analysis and sequenced to confirm thecorrect insert.

EXAMPLE 55

[1133] Construction of pMON710

[1134] Plasmid pMON709 consists of a 1614 base pair AvaI,EcoRI fragmentof transposon TN7, containing the streptomycin adenylyltransferase gene(Fling et al., 1985) and a pUC9 linker (XmaI,HindIII) cloned between theHindIII and EcoRI sites of pUC19. The streptomycin adenylyltransferasegene COnfers resistance to streptomycin and spectinomycin. PlasmidpMON709 was mutagenized by oligonucleotide site-directed mutagenesis(methods described in Zoller and Smith, 1982) to introduce an EcoRV siteat the 3′ end of the streptomycin adenylyltransferase gene. Theoligonucleotide, Oligo # 53 [SEQ ID NO:157], was used in this procedureto introduce the EcoRV site. The resulting plasmid was designatedpMON710.

EXAMPLE 56

[1135] Construction of pMON5723

[1136] Plasmid pMON5647 DNA was digested with restriction enzymes DraIand SspI resulting in a 2916 base pair DraI, SspI fragment. The geneticelements derived from pMON5647 are the pBR327 origin of replication,precA promoter, g10L ribosome binding protein, porcine somatotropin gene(pST) and T7 transcription terminator (Dunn and Strudier, 1983). PlasmidpMON710 DNA was digested with restriction enzymes HincII and EcoRVresulting in 940 base pair HincII,EcoRV fragment containing thestreptomycin adenylyltransferase gene which infers resistance tostreptomycin and spectinomycin. The restriction fragments were ligatedand the ligation reaction mixture was used to transform E. coli strainJM101. The DraI, SspI, HincII and EcoRV restriction sites are lost as aresult of the cloning. Transformant bacteria were selected onspectinomycin-containing plates. Plasmid DNA was isolated, analyzed byrestriction analysis and sequenced to confirm the correct insert.

EXAMPLE 57

[1137] Construction of pMON13361

[1138] Plasmid pMON13288 was mutagenized by oligonucleotidesite-directed mutagenesis (method described in Kunkel, 1985) toeliminate a NsiI site in the (15-125) hIL-3 variant coding region. Codon70 of (15-125) hIL-3, encoding asparagine, was converted from AAT to AACdestroying the NsiI recognition site. The oligonucleotide, Oligo # 54[SEQ ID NO:158], was used in this procedure to eliminate the NsiI site.Transformant bacteria were selected on ampicillin-containing plates.Plasmid DNA was isolated, analyzed by restriction analysis to confirmthe loss of the NsiI site and sequenced to confirm the sequence of the(15-125) hIL-3 variant gene. The plasmid, pMON13361, encodes the(15-125) hIL-3 variant with the amino acid sequence of PEPTIDE #25 [SEQID NO:89].

[1139] DNA sequence # 32 [SEQ ID NO:160] codes for the foregoingpMON13361 polypeptide.

EXAMPLE 58

[1140] Construction of pMON14058

[1141] Plasmid pMON13361 was mutagenized by oligonucleotidesite-directed mutagenesis (method described by Taylor et al., 1985 usinga kit from Amersham, Arlington Heights, Ill.) to eliminate a EcoRV sitein the (15-125) hIL-3 variant coding region. Codon 46 and 47 of (15-125)hIL-3, encoding asparagine and isoleucine, were converted from GAT toGAC and ATC to ATT respectively, destroying the EcoRV recognition site.The oligonucleotide, Oligo # 55 [SEQ ID NO:159], was used in thisprocedure to eliminate the EcoRV site. Transformant bacteria wereselected on ampicillin-containing plates. Plasmid DNA was isolated,analyzed by restriction analysis to confirm the loss of the EcoRV siteand sequenced to confirm the sequence of the (15-125) hIL-3 variantgene. The plasmid, pMON14058, encodes the (15-125) hIL-3 variant withthe amino acid sequence of PEPTIDE #25 [SEQ ID NO:89].

[1142] DNA sequence # 33 [SEQ ID NO:161] codes for the foregoingpMON14058 polypeptide.

EXAMPLE 59

[1143] Construction of pMON13438

[1144] Plasmid pMON5723 DNA was digested with restriction enzymes NcoIand HindIII resulting in a 3278 NcoI,HindIII fragment. The geneticelements derived from pMON5723 are the pBR327 origin of replication,precA promoter, g10L ribosome binding protein, T7 transcriptionterminator and streptomycin adenylyltransferase gene. Plasmid pMON14058DNA was digested with NcoI and HindIII resulting in a 345 base pairNcoI,HindIII fragment which contains the (15-125) hIL-3 gene with thefollowing amino acid substitutions: 18I, 25H, 29R, 32N, 37P, 42S, 45M,51R, 55T, 59L, 62V, 67H, 69E,73G, 76A, 79R, 83Q, 87S, 93S, 98I, 101A,105Q, 109E, 116V, 120Q and 123E. The restriction fragments were ligatedand the ligation reaction mixture was used to transform E. coli strainJM101. Transformant bacteria were selected on spectinomycin-containingplates. Plasmid DNA was isolated, analyzed by restriction analysis andsequenced to confirm the correct insert. The plasmid, pMON13438, encodesthe (15-125) hIL-3 variant with the amino acid sequence of PEPTIDE #25[SEQ ID NO:89].

[1145] DNA sequence # 33 [SEQ ID NO:161] codes for the foregoingpMON13438 polypeptide.

EXAMPLE 60

[1146] Construction of pMON13285

[1147] Plasmid pMON13252 DNA was digested with restriction enzymes NcoIand EcoRV and the resulting 3669 base pair NcoI,EcoRV fragment containsthe following genetic elements; streptomycin adenyltransferase gene,pBR327 origin of replication, phage f1 origin of replication as thetranscription terminator, recA promoter, g10L ribosome binding site andthe bases encoding amino acids 47-125 of (15-125) hIL-3 with thefollowing amino acid substitution, 50D. The 3669 base pair NcoI,EcoRVrestriction fragment from pMON13252 was ligated to the followingannealed complementary oligonucleotides.

[1148] Oligo #165 [SEQ ID NO:162]

[1149] Oligo #166 [SEQ ID NO:163]

[1150] Oligo #167 [SEQ ID NO:164]

[1151] Oligo #168 [SEQ ID NO:165]

[1152] Oligo #169 [SEQ ID NO:166]

[1153] Oligo #170 [SEQ ID NO:167]

[1154] When assembled, the oligonucleotides create NcoI and EcoRVrestriction ends and the DNA sequence that encodes amino acids 15-46 of(15-125) hIL-3 with the following amino acid substitutions; 42D, 45M and46S. The codons encoding amino acids 15-46 of (15-125) hIL-3 are thosefound in the hIL-3 cDNA sequence except at those positions where aminoacid substitutions were made. The plasmid, pMON13285, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#A3 [SEQ ID NO:258] DNA sequence #A3 pMON13285 42D, 45M 46S, 50DATGGCTAACT GCTCTAACAT GATCGATGAA [SEQ ID NO:398] ATCATCACCC ACCTGAAGCAGCCACCGCTC CCCCTGCTGG ACTTCAACAA CCTCAATGAC GAAGACATGT CTATCCTGATGGACAATAAC CTTCGTCGTC CAAACCTCGA GCCATTCAAC CGTGCTGTCA ACTCTCTCCAGAATGCATCA GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCACGGCCGCACCC ACCCCACATC CAATCCATAT CAACGACCGT GACTGGAATG AATTCCGTCGTAAACTGACC TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG

EXAMPLE 61

[1155] Construction of pMON13286

[1156] Plasmid pMON5978 DNA was digested with restriction enzymes NcoIand EcoRV and the resulting 3865 base pair NcoI,EcoRV fragment containsthe following genetic elements; beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, precA promoter, g10L ribosome binding site and the basesencoding amino acids 47-125 of (15-125) hIL-3. The 3865 base pairNcoI,EcoRV restriction fragment from pMON5978 was ligated to thefollowing annealed complementary oligonucleotides.

[1157] Oligo #165 [SEQ ID NO:162]

[1158] Oligo #166 [SEQ ID NO:163]

[1159] Oligo #167 [SEQ ID NO:164]

[1160] Oligo #168 [SEQ ID NO:165]

[1161] Oligo #169 [SEQ ID NO:166]

[1162] Oligo #170 [SEQ ID NO:167]

[1163] When assembled, the oligonucleotides create NcoI and EcoRVrestriction ends and the DNA sequence that encodes amino acids 15-46 of(15-125) hIL-3 with the following amino acid substitutions; 42D, 45M and46S. The codons encoding amino acids 15-46 of (15-125) hIL-3 are thosefound in the hIL-3 cDNA sequence except at those positions where aminoacid substitutions were made. The plasmid, pMON13286, encodes the(15-125) hIL-3 variant with the following amino acid sequence: Peptide#A4 [SEQ ID NO:259] DNA sequence #A4 pMON13286 42D, 45M, 46S ATGGCTAACTGCTCTAACAT GATCGATGAA [SEQ ID NO:399] ATCATCACCC ACCTGAAGCA GCCACCGCTGCCGCTGCTGG ACTTCAACAA CCTCAATGAC GAAGACATGT CTATCCTGAT GGAAAATAACCTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCA AGTCTCTGCA GAATGCATCAGCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGC CCCTGGCCAC GGCCGCACCCACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATG AATTCCGTCG TAAACTGACCTTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG

EXAMPLE 62

[1164] Construction of pMON13325

[1165] The 3704 base pair EcoRI, HindIII DNA fragment from plasmidpMON13286 is ligated to the 64 base pair EcoRI, HindIII DNA fragmentfrom plasmid pMON13215. The following genetic elements are derived frompMON13286; beta-lactamase gene (AMP), pBR327 origin of replication,phage F1 origin of replication as the transcription terminator, precApromoter, g10L ribosome binding site and the bases encoding amino acids15-105 of the (15-125) hIL-3 gene with the following changes, 42D, 45M,and 46S. The bases encoding amino acids 106-125 of the (15-125) genewith the following change, 116W, are derived from pMON13215. Theresulting plasmid, pMON13325, encodes the (15-125) hIL-3 variant withthe following amino acid sequence:

[1166] Peptide # A5 [SEQ ID NO:261]

EXAMPLE 63

[1167] Construction of pMON13326

[1168] The 3683 base pair NcoI, EcoRI DNA fragment from plasmidpMON13215 is ligated to the 281 base pair NcoI, EcoRI DNA fragment fromplasmid pMON13285. The following genetic elements are derived frompMON13215; beta-lactamase gene (AMP), pBR327 origin of replication,phage F1 origin of replication as the transcription terminator, precApromoter, g10L ribosome binding site and the bases encoding amino acids106-125 of the (15-125) hIL-3 gene with the following change, 116W. Thebases encoding amino acids 15-105 of the (15-125) gene with thefollowing change, 42D, 45M, 46S and 50D derived from pMON13285. Theresulting plasmid, pMON13326, encodes the (15-125) hIL-3 variant withthe following amino acid sequence:

[1169] Peptide # A6 [SEQ ID NO:262]

EXAMPLE 64

[1170] Construction of pMON13332

[1171] Plasmid pMON13326 DNA is digested with restriction enzymes NsiIand EcoRI and the resulting 3853 base pair NsiI,EcoRI fragment containsthe following genetic elements; beta-lactamase gene (AMP), pBR327 originof replication, phage f1 origin of replication as the transcriptionterminator, recA promoter, g10L ribosome binding site and the basesencoding amino acids 15-71 and 106-125 of (15-125) hIL-3 gene with thefollowing changes 42D, 45M, 46S, 50D and 116W. The 3853 base pairNsiI,EcoRI restriction fragment from pMON13326 is ligated to thefollowing annealed complementary oligonucleotides.

[1172] Oligo #15(A) [SEQ ID NO:168]

[1173] Oligo #16(A) [SEQ TD NO:169]

[1174] In the resulting plasmid the 111 bases between the NsiI and EcoRIrestriction sites in the (15-125) hIL-3 gene are replaced with 24 basesfrom the above mentioned oligonucleotides. This linker also creates aNdeI recognition sequence.

EXAMPLE 65

[1175] Construction of pMON13330

[1176] The 3846 base pair PstI, EcoRI DNA fragment from plasmidpMON13332 is ligated to the 118 base pair PstI, EcoRI DNA fragment fromplasmid pMON13305. The following genetic elements are derived frompMON13332; beta-lactamase gene (AMP), pBR327 origin of replication,phage f1 origin of replication as the transcription terminator, recApromoter, g10L ribosome binding site and the bases encoding amino acids15-69 and 106-125 of the (15-125) hIL-3 gene with the following change,42D, 45M, 46S, 50D and 116W. The bases encoding amino acids 70-105 ofthe (15-125) gene with the following change, 95R, 98T and 100R arederived from pMON13305. The resulting plasmid, pMON13330, encodes the(15-125) hIL-3 variant with the following amino acid sequence:

[1177] Peptide # A7 [SEQ ID NO:263]

EXAMPLE 66

[1178] Construction of pMON13329

[1179] The 3846 base pair PstI, EcoRI DNA fragment from plasmidpMON13332 is ligated to the 118 base pair PstI, EcoRI DNA fragment fromplasmid pMON13304. The following genetic elements are derived frompMON13332; beta-lactamase gene (AMP), pBR327 origin of replication,phage f1 origin of replication as the transcription terminator, recApromoter, g10L ribosome binding site and the bases encoding amino acids15-69 and 106-125 of the (15-125) hIL-3 gene with the following change,42D, 45M, 46S, and 116W. The bases encoding amino acids 70-105 of the(15-125) gene with the following change, 98I and 100R are derived frompMON13304. The resulting plasmid, pMON13329, encodes the (15-125) hIL-3variant with the following amino acid sequence:

[1180] Peptide # A8 [SEQ ID NO:406]

EXAMPLE 67

[1181] Construction of pMON5853 (FIG. 6) which encodes [Met-(15-133)hIL-3(Arg¹²⁹)]

[1182] Plasmid DNA of pMON5847 (Example 2) was treated with NcoI. Therestriction enzyme was inactivated by heat treatment (65° C. for 10minutes). The DNA was then treated with large fragment of DNA polymeraseI (Klenow) in the presence of all four nucleotide precursors. Thisproduces DNA termini with non-overlapping ends. After 5 minutes at 37°C., the polymerase was inactivated by heat treatment at 65° C. for 10minutes. The DNA was then treated with HpaI, an enzyme which producesnon-overlapping termini. The DNA was ethanol precipitated and ligated.The ligation reaction mixture was used to transform competent JM101cells to ampicillin resistance. Colonies were picked and plasmid DNA wasanalyzed by restriction analysis. A plasmid designated pMON5853 wasidentified as one containing a deletion of the amino terminal 14 codonsof the hIL-3 gene. The DNA sequence for the junction of the ribosomebinding site to the (15-133) hIL-3 gene was determined to be thefollowing: [SEQ ID NO:400] [SEQ ID NO:401]5′-AAGGAGATATATCCATGAACTGCTCTAAC-3′                  M  N  C  S  N

[1183] The lower line contains the one-letter code for the amino acidsspecified by the coding sequence of the amino terminus of the 15-133hIL-3 gene. These are methionine, asparagine, cysteine, serine andasparagine.

[1184] When cultures of JM101 cells harboring this plasmid were inducedwith nalidixic acid, it was found that hIL-3 (15-133) accumulated atlevels higher than hIL-3 (pMON5847).

[1185] The plasmid, pMON5853, encodes Met-(15-133) hIL-3 (Arg¹²⁹) whichhas the following amino acid sequence:     Met Asn Cys Ser Asn Met IleAsp Glu Ile Ile Thr [SEQ ID NO:402] His Leu Lys Gln Pro Pro Leu Pro LeuLeu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn AsnLeu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln AsnAla Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu AlaThr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn GluPhe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala GlnGln Thr Thr Leu Arg Leu Ala Ile Phe

EXAMPLE 68

[1186] Construction of pMON13252

[1187] Plasmid, pMON2341, DNA was digested with restriction enzymes NcoIand HindIII resulting in a 3619 base pair NcoI/HindIII fragment. Thegenetic elements derived from pMON2341 are the beta-lactamase gene(AMP), pBR327 origin of replication F1 phage origin of replication asthe transcription terminator, precA, g10L ribosome binding site. Theplasmid encoding the hIL-3 (15-125) Asp⁽⁵⁰⁾ variant, was digested withNcoI and HindIII resulting in a 345 base pair NcoI/HindIII fragment.This 345 Base pair NcoI/HindIII fragment was ligated with the 3619 basepair fragment from pMON2341 and the ligation reaction mixture was usedto transform E. coli K-12 strain JM101. Plasmid DNA was isolated andscreened by restriction analysis using NcoI and HindIII. Positive clonescontained a 345 base pair NcoI/HindIII fragment. This construct wasdesignated pMON13252. The plasmid, pMON13252, encodes the (15-125) hIL-3variant with the following amino acid sequence: PEPTIDE A10;(15-125)HIL-3 Asp⁽⁵⁰⁾ pMON13252         Asn Cys Ser Asn Met Ile Asp GluIle Ile Thr His Leu [SEQ ID NO:407]        15                  20                  25 Lys Gln Pro Pro LeuPro Leu Leu Asp Phe Asn Asn Leu Asn Gly        30                  35                  40 Glu Asp Gln Asp IleLeu Met Asp Asn Asn Leu Arg Arg Pro Asn        45                  50                  55 Leu Glu Ala Phe AsnArg Ala Val Lys Ser Leu Gln Asn Ala Ser        60                  65                  70 Ala Ile Glu Ser IleLeu Lys Asn Leu Leu Pro Cys Leu Pro Leu        75                  80                  85 Ala Thr Ala Ala ProThr Arg His Pro Ile His Ile Lys Asp Gly        90                  95                  100 Asp Trp Asn Glu PheArg Arg Lys Leu Thr Phe Tyr Leu Lys Thr        105                 110                 115 Leu Glu Asn Ala GlnAla Gln Gln         120                 125 DNA sequence #A10 pMON1325250D ATGGCTAACT GCTCTAACAT GATCGATGAA ATCATCACCC ACCTGAAGCA [SEQ IDNO:408] GCCACCGCTG CCGCTGCTGG ACTTCAACAA CCTCAATGGT GAAGACCAAGATATCCTGAT GGAACAATAAC CTTCGTCGTC CAAACCTCGA GGCATTCAAC CGTGCTGTCAACTCTCTGCA GAATGCATCA GCAATTGAGA GCATTCTTAA AAATCTCCTG CCATGTCTGCCCCTGGCCAC GGCCGCACCC ACGCGACATC CAATCCATAT CAAGGACGGT GACTGGAATGAATTCCGTCG TAAACTGACC TTCTATCTGA AAACCTTGGA GAACGCGCAG GCTCAACAG

EXAMPLES 69-76

[1188] The variants in Table 5 were constructed by cassette mutagenesisusing methods described in the Materials and Methods and the Examplescontained herein, particularly Examples 54-58. Parental plasmid DNA(Table 5), digested with the appropriate restriction enzymes (Table 5),was ligated with the indicated annealed pairs of complementaryoligonucleotides (Table 5). The assembled oligonucleotides createappropriate restriction ends and a portion of the (15-125) hIL-3 genesequence (pMON13288 [SEQ ID NO:100]). Individual isolates were screenedby restriction analysis and DNA sequenced to confirm that the desiredchanges in the (15-125) hIL-3 variant gene were made. Theoligonucleotides create change(s) in the (15-125) hIL-3 gene whichencode the corresponding amino acid substitution(s) in the variantpolypeptide (Table 5). The amino acids substitutions in addition toand/or different from those in polypeptide # 25 [SEQ ID NO:89] areindicated in Table 5. The table also shows the plasmid designation (pMONnumber), DNA sequence identification number for the mutated hIL-3 geneand the identification number for the the resulting variant polypeptide.The biological activity (growth promoting activity in AML 193 cells) forsome of the variants in Table 5 is shown in Table 1.

EXAMPLES 77-82

[1189] The variants in Table 6 were constructed by methods described inthe Materials and Methods and the Examples contained herein,particularly in Examples 60 and 61. Parental plasmid DNA (Table 6),digested with the appropriate restriction enzymes (Table 6), was ligatedwith the indicated restriction fragment (Table 6). Individual isolateswere screened by restriction analysis and DNA sequenced to confirm thatthe desired changes in the (15-125) hIL-3 variant gene were made. Theresulting mutated (15-125) hIL-3 genes encode the corresponding aminoacid substitutions in the variant polypeptides (Table 6). The aminoacids substitutions in addition to and/or different from those inpolypeptide # 25 [SEQ ID NO:89] are indicated in Table 6. The table alsoshows the plasmid designation (pMON number), DNA sequence identificationnumber for the mutated hIL-3 gene and the identification number for thethe resulting variant polypeptide. The biological activity (growthpromoting activity in AML 193 cells) for some of the variants in Table 6is shown in Table 1.

EXAMPLE 83

[1190] Construction of pMON13368

[1191] One of the DNA fragments to construct the plasmid, pMON13368, wasgenerated by site-directed mutagenesis employing PCR techniquesdescribed in the Materials and Methods and the Examples containedherein, particularly Example 53. The template for the PCR reaction wasplasmid, pMON13289, DNA using the oligonucleotides, Oligo #B13 18123A25H[SEQ ID NO: 182] and Oligo #B14 2341HIN3 [SEQ ID NO:183], as primers.The resulting DNA product was digested with the restriction enzymes NcoIand HindIII. Upon completion, the digest was heated at 70□C for 15minutes to inactivate the enzymes. The restriction fragment was purifiedby phenol/chloroform extraction and precipitation with equal volumeisopropanol in the presence of 2M NH4OAc. The oligonucleotide, Oligo#B13 18I23A25H [SEQ ID NO:182], changes the codon at position 23 of(15-125) hIL-3 variant gene pMON13289 [SEQ ID NO:103] from ‘ATT’ to‘GCA’ (Ile to Ala). The 3619 base pair NcoI, HindIII restrictionfragment from pMON2341 was ligated to the PCR-generated NcoI, HindIIIrestriction fragment. Individual isolates were screened by restrictionanalysis and DNA sequenced to confirm that the desired changes in the(15-125) hIL-3 variant gene were made. The plasmid, pMON13368, containsthe (15-125) hIL-3 variant gene (DNA sequence #B15 [SEQ ID NO:346])which encodes the (15-125) hIL-3 variant polypeptide with the followingamino acid sequence:

[1192] Polypeptide #B15 [SEQ ID NO.:278]

EXAMPLE 84

[1193] Construction of pMON13380

[1194] Plasmid, pMON13368, DNA was digested with restriction enzymesEcoRI and HindIII. The resulting 3900 base pair EcoRI,HindIII fragmentcontains the following genetic elements; beta-lactamase gene (AMP),pBR327 origin of replication, phage F1 origin of replication as thetranscription terminator, precA promoter, g10L ribosome binding site andthe DNA sequence encoding amino acids 15-105 of the variant pMON13368.The 3900 base pair EcoRI,HindIII restriction fragment from pMON13368 wasligated to the following annealed complementary oligonucleotides. Oligo# B48 9E12Q6V1 [SEQ ID NO: 217] Oligo # B49 9E12Q6V3 [SEQ ID NO: 218]Oligo # 49 120Q123E2 [SEQ ID NO: 63] Oligo # 50 120Q123E4 [SEQ ID NO:64]

[1195] When assembled, the oligonucleotides create EcoRI and HindIIIrestriction ends and the DNA sequence that encodes amino acids 106-125of (15-125) hIL-3 with the following amino acid substitution; 109E,112Q, 116V, 120Q and 123E. The codons used in the (15-125) hIL-3 geneare those found in the hIL-3 cDNA sequence except at those positionswhere amino acid substitutions were made. Individual isolates werescreened by restriction analysis and DNA sequenced to confirm that thedesired changes in the (15-125) hIL-3 variant gene were made. Theplasmid, pMON13380, contains the (15-125) hIL-3 variant gene (DNAsequence #B16 [SEQ ID NO:347]) which encodes the (15-125) hIL-3 variantpolypeptide with the following amino acid sequence:

[1196] Polypeptide #B16 [SEQ ID NO.:279]

EXAMPLE 85

[1197] Construction of pMON13476

[1198] One of the DNA fragments to construct the plasmid, pMON13476, wasgenerated by site-directed mutagenesis employing PCR techniquesdescribed in the Materials and Methods and the Examples containedherein, particularly Example 54. The template for the PCR reaction wasplasmid, pMON13287, DNA using the oligonucleotides, Oligo #B13 18I23A25H[SEQ ID NO:182] and Oligo #B14 2341HIN3 [SEQ ID NO.:183] as primers. Theresulting DNA product was digested with the restriction enzymes NcoI andHindIII. Upon completion, the digest was heated at 70□C for 15 minutesto inactivate the enzymes. The restriction fragment was purified byphenol/chloroform extraction and precipitation with equal volumeisopropanol in the presence of 2M NH4OAc. The oligonucleotide, Oligo#B13 18I23A25H [SEQ ID NO.:182], changes the codon at position 23 of(15-125) hIL-3 variant gene, pMON13287, [SEQ ID NO:97] from ‘ATT’ to‘GCA’ (Ile to Ala). The 3619 base pair NcoI, HindIII restrictionfragment from pMON2341 was ligated to the PCR-generated NcoI, HindIIIrestriction fragment. Individual isolates were screened by restrictionanalysis and DNA sequenced to confirm that the desired changes in the(15-125) hIL-3 variant gene were made. The resulting clone alsocontained a change, that was not designed in the mutagenicoligonucleotide, which changed the codon at position -1 from ‘GCT’ to‘GAT’ which changes the amino acid from Alanine to Aspartic Acid. Theplasmid, pMON13476, contains the (15-125) hIL-3 variant gene (DNAsequence #B52 [SEQ ID NO:303]) which encodes the (15-125) hIL-3 variantpolypeptide with the following amino acid sequence:

[1199] Polypeptide #B52 [SEQ ID NO.:314]

EXAMPLES 86-92

[1200] The variants in Table 7 were constructed by PCR techniques usingmethods described in the Materials and Methods and the Example containedherein, particularly Example 51. Two sequential PCR reactions were usedto create the variants. In the first PCR reaction pMON13287 plasmid DNAserved as the template and the two oligonucleotides indicated in Table 7served as the primers. Following the PCR extension reaction, the PCRproduct was partially purified to remove primer that was not extended.In the second PCR reaction pMON13287 plasmid DNA served as the template,the purified PCR product from the first PCR reaction served as one ofthe primers and the Oligo #B14 2341Hin3 [SEQ ID NO:183] as the secondprimer. The product from the second PCR reaction was partially purifiedand digested with restriction enzymes NcoI and HindIII and ligated withthe 3619 base pair NcoI,HindIII fragment from pMON2341. Individualisolates were screened by restriction analysis and DNA sequenced toconfirm that the desired changes in the (15-125) hIL-3 variant gene weremade. The amino acids substitutions in addition to and/or different fromthose in polypeptide # 24 [SEQ ID NO:88] are indicated in Table 7. Thetable also shows the plasmid designation (pMON number), DNA sequenceidentification number for the mutated hIL-3 gene and the identificationnumber for the the resulting variant polypeptide. The biologicalactivity (growth promoting activity in AML 193 cells) for some of thevariants in Table 7 is shown in Table 1.

EXAMPLES 93-120

[1201] The variants in Table 8 were constructed by cassette mutagenesisusing methods described in the Materials and Methods and the Examplescontained here, particularly Examples 54-58. Parental plasmid DNA (Table8), digested with the appropriate restriction enzymes (Table 8), wasligated with the indicated annealed pairs of complementaryoligonucleotides (Table 8). The assembled oligonucleotides create theappropriate restriction ends and a portion of (15-125) hIL-3 gene(pMON13288 [SEQ ID NO:100]) sequence. The oligonucleotides createchange(s) in the (15-125) hIL-3 variant gene which encode thecorresponding amino acid substitution(s); and/or deletions from theC-terminus of the variant polypeptide (Table 8). Individual isolateswere screened by restriction analysis and DNA sequenced to confirm thatthe desired changes in the (15-125) hIL-3 variant gene were made. Theamino acids substitutions in addition to and/or different from those inpolypeptide # 25 [SEQ ID NO:88] are indicated in Table 8. The table alsoshows the plasmid designation (pMON number), DNA sequence identificationnumber for the mutated hIL-3 gene and the identification number for thethe resulting variant polypeptide. The biological activity (growthpromoting activity in AML 193 cells) for some of the variants in Table 5is shown in Table 1.

EXAMPLE 121

[1202] Construction of pMON13446

[1203] Plasmid, pMON13287, DNA (purified from the E. coli strain GM48{dam-}) was digested with restriction enzymes NcoI and ClaI. Theresulting 3942 base pair NcoI,ClaI fragment contains the followinggenetic elements; beta-lactamase gene (AMP), pBR327 origin ofreplication, phage F1 origin of replication as the transcriptionterminator, precA promoter, g10L ribosome binding site and the DNAsequence encoding amino acids 21-125 of the (15-125) hIL-3 variantpMON13287. The 3942 base pair NcoI,ClaI restriction fragment frompMON13368 was ligated to the following annealed complementaryoligonucleotides. Oligo # B57 338UP [SEQ ID NO: 226] Oligo # B56 338DOWN[SEQ ID NO: 225]

[1204] When assembled, the oligonucleotides create NcoI and ClaIrestriction ends and the DNA sequence that encodes the following 14amino acid sequence; Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp AspLys [SEQ ID NO:403] and the DNA sequence which encodes amino acids 15-20of the (15-125) hIL-3 variant gene, pMON13287 [SEQ ID NO:97]. Theresulting variant polypeptide has a 14 amino acid N-terminal extensionfused to the (15-125) hIL-3 variant polypeptide, pMON13288 [SEQ ID NO:88]. The plasmid, pMON13446, contains the (15-125) hIL-3 variant gene(DNA sequence #B53 [SEQ ID NO:404]) which encodes the (15-125) hIL-3variant polypeptide with the following amino acid sequence:

[1205] Polypeptide #B53 [SEQ ID NO.:315]

EXAMPLE B54

[1206] Construction of pMON13390

[1207] Plasmid, pMON13288, DNA (purified from the E. coli strain GM48{dam-}) was digested with restriction enzymes NcoI and ClaI. Theresulting 3942 base pair NcoI,ClaI fragment contains the followinggenetic elements; beta-lactamase gene (AMP), pBR327 origin ofreplication, phage F1 origin of replication as the transcriptionterminator, precA promoter, g10L ribosome binding site and the DNAsequence encoding amino acids 21-125 of the (15-125) hIL-3 variantpMON13288. The 3942 base pair NcoI,ClaI restriction fragment frompMON13288 was ligated to the following annealed complementaryoligonucleotides. Oligo # B57 338UP [SEQ ID NO: 226] Oligo # B56 338DOWN[SEQ ID NO: 225]

[1208] When assembled, the oligonucleotides create NcoI and ClaIrestriction ends and the DNA sequence which encodes the following 14amino acid sequence; Met Ala Tyr Pro Glu Thr Asp Tyr Lys Asp Asp Asp AspLys [SEQ ID NO:403] and the DNA sequence which encodes amino acids 15-20of the (15-125) hIL-3 variant gene pMON13288 [SEQ ID NO:100]. Theresulting variant has a 14 amino acid N-terminal extension fused to the(15-125) hIL-3 variant polypeptide, pMON13288 [SEQ ID NO:88]. Theplasmid, pMON13390, containes the (15-125) hIL-3 variant gene (DNAsequence #B54 [SEQ ID NO.:405] which encodes the (15-125) hIL-3 variantpolypeptide with the following amino acid sequence:

[1209] Polypeptide #B54 [SEQ ID NO:316]

EXAMPLES 133-136

[1210] The variants in Table 10 were constructed by methods described inMaterials and Methods and in Examples contained herein, particularlyExamples 54-58. Parental plasmid DNA (Table 10), digested with theappropriate restriction enzymes (Table 10) was ligated with theindicated restriction fragment containing the changes listed (Table 10).The resulting mutated (15-125) IL-3 genes encode the corresponding aminoacid substitutions in the variant polypeptides (Table 10). The aminoacid substitutions in addition to and/or different from those inpolypeptide #25 [SEQ ID NO: 89] are indicated in Table 10. Thebiological activity (growth promoting activity in AML 193 cells) forsome of the variants in Table 10 is shown in Table 1.

EXAMPLES 123-132

[1211] The variants in Table 9 were constructed by cassett mutagenesisusing methods described in Materials and Methods and in Examples 54-58contained herein. Parental plasmid DNA (Table 9), digested with theappropriate restriction enzymes (Table 9), was ligated with theindicated annealed pairs of complementry oligonucleoties (Table 9). Theassembled oligonucleotides create the appropriate restriction fragmentwhich was inserted into the (15-125) hIL-3 gene (pMON13288 [SEQ IDNO:100] between these restriction sites. The deletions or substitutionsencoded by the oligonucleotide in the (15-125) IL-3 gene correspond tothe amino acid deletions or substitutions in the variant polypeptide(Table 9). The amino acid substitutions or deletions, in addition toand/or different from those in the polypeptide #25 [SEQ ID NO:89] areindicated in Table 9. The biological activity (growth promoting activityin AML 193 cells) for some of the variants in Table 9 is shown in Table1.

[1212] Formula XI shown below is a representation of a [(15-125) hIL-3mutein] with numbers in bold type added above the amino acids torepresent the position at which the amino acid below the bolded numberappears in native (1-133) hIL-3 [e. g. the amino acid at position 1 ofFormula XI corresponds to the Asn which appears at position 15 in native(1-133) hIL-3]. The number shown in bold indicates the amino acids thatcorrespond to the native IL-3(1-133). The non-bold members below theamino acids sequences are for Seq Id reference numbers. When the muteinsare expressed the initial amino acid may be preceded by Met- orMet-Ala-.  15              20                25 Asn Cys Ser Asn Met IleAsp Glu Ile Ile Thr His Leu Lys Gln [SEQ ID NO:23] 1                5                  10                  15 30              35                40 Pro Pro Leu Pro Leu Leu Asp PheAsn Asn Leu Asn Gly Glu Asp                 20                  25                  30 45              50                55 Gln Asp Ile Leu Met Glu Asn AsnLeu Arg Arg Pro Asn Leu Glu                 35                  40                  45 60              65                70 Ala Phe Asn Arg Ala Val Lys SerLeu Gln Asn Ala Ser Ala Ile                 50                  55                  60 75              80                85 Glu Ser Ile Leu Lys Asn Leu LeuPro Cys Leu Pro Leu Ala Thr                 65                  70                  75 90              95               100 Ala Ala Pro Thr Arg His Pro IleHis Ile Lys Asp Gly Asp Trp                 80                  85                  90105              110               115 Asn Glu Phe Arg Arg Lys Leu ThrPhe Tyr Leu Lys Thr Leu Glu                 95                 100                 105120              125 Asn Ala Gln Ala Gln Gln                 110

[1213] TABLE 5 Parental plasmid/ amino acid Example pMON numberrestriction digest oligo pair 1, 4 oligo pair 2, 5 oligo pair 3, 6changes resulting polypeptide Example 69 pMON13406 pMON13288/ 19Ala129R32N37P2 42S45M3 19Ala polypeptide B1 SEQ ID NO: 332 NcoI, EcoRVOLIGO# B1 OLIGO# 5 OLIGO# 11 SEQ ID NO: 264 SEQ ID NO: 170 SEQ ID NO: 19SEQ ID NO: 25 19Ala4 29R32N37P5 42S45M6 OLIGO# B2 OLIGO# 6 OLIGO# 12 SEQID NO: 171 SEQ ID NO: 20 SEQ ID NO: 26 Example 70 pMON13414 pMON13288/19Ile1 29R32N37P2 42S45M3 19Ile polypeptide B2 SEQ ID NO: 333 NcoI,EcoRV OLIGO# B3 OLIGO# 5 OLIGO# 11 SEQ ID NO: 265 SEQ ID NO: 172 SEQ IDNO: 19 SEQ ID NO: 25 19Ile4 29R32N37P5 42S45M6 OLIGO# B4 OLIGO# 6 OLIGO#12 SEQ ID NO: 173 SEQ ID NO: 20 SEQ ID NO: 26 Example 71 pMON13407pMON13288/ 18I25H1 29R32N37P2 42S45V3 45Val polypeptide B3 SEQ ID NO:334 NcoI, EcoRV OLIGO# 1 OLIGO# 5 OLIGO# B11 SEQ ID NO: 266 SEQ ID NO:15 SEQ ID NO: 19 SEQ ID NO: 180 18I25H4 29R32N37P5 42S45V6 OLIGO# 2OLIGO# 6 OLIGO# B12 SEQ ID NO: 16 SEQ ID NO: 20 SEQ ID NO: 181 Example72 pMON13405 pMON13288/ 19Ala1 29R32N37P2 42S45V3 19Ala, 45Valpolypeptide B4 SEQ ID NO: 335 NcoI, EcoRV OLIGO# B1 OLIGO# 5 OLIGO# B11SEQ ID NO: 267 SEQ ID NO: 170 SEQ ID NO: 19 SEQ ID NO: 180 19Ala429R32N37P5 42S45V6 OLIGO# B2 OLIGO# 6 OLIGO# B12 SEQ ID NO: 171 SEQ IDNO: 20 SEQ ID NO: 181 Example 73 pMON13415 pMON13288/ 19Ile1 29R32N37P242S45V3 19Ile, 45Val polypeptide B5 SEQ ID NO: 336 NcoI, EcoRV OLIGO# B3OLIGO# 5 OLIGO# B11 SEQ ID NO: 268 SEQ ID NO: 172 SEQ ID NO: 19 SEQ IDNO: 180 19Ile4 29R32N37P5 42S45V6 OLIGO# B4 OLIGO# 6 OLIGO# B12 SEQ IDNO: 173 SEQ ID NO: 20 SEQ ID NO: 181 Example 74 pMON13408 pMON13288/49Ile1 59L62V2 67H69E3 49Ile polypeptide B6 SEQ ID NO: 337 EcoRV, NsiIOLIGO# B7 OLIGO# 25 OLIGO# 29 SEQ ID NO: 269 SEQ ID NO: 176 SEQ ID NO:39 SEQ ID NO: 43 49Ile4 59L62V5 67H69E6 OLIGO# B8 OLIGO# 26 OLIGO# 30SEQ ID NO: 177 SEQ ID NO: 40 SEQ ID NO: 44 Example 75 pMON13409pMON13288/ 49Leu1 59L62V2 67H69E3 49Leu polypeptide B7 SEQ ID NO: 338EcoRV, NsiI SEQ ID NO: 178 OLIGO# 25 OLIGO# 29 SEQ ID NO: 270 OLIGO# B9SEQ ID NO: 39 SEQ ID NO: 43 49Leu4 59L62V5 67H69E6 OLIGO# B10 OLIGO# 26OLIGO# 30 SEQ ID NO: 179 SEQ ID NO: 40 SEQ ID NO: 44 Example 76pMON13410 pMON13288/ 49Asp1 59L62V2 67H69E3 49Asp polypeptide B8 SEQ IDNO: 339 EcoRV, NsiI OLIGO# B5 OLIGO# 25 OLIGO# 29 SEQ ID NO: 271 SEQ IDNO: 174 SEQ ID NO: 39 SEQ ID NO: 43 49Asp4 59L62V5 67H69E6 OLIGO# B6OLIGO# 26 OLIGO# 30 SEQ ID NO: 175 SEQ ID NO: 40 SEQ ID NO: 44

[1214] TABLE 6 plasmid pMON Parental plasmid/ restriction amino acidresulting Example No number restriction digest fragment substitutionspolypeptide Example 77 pMON13422 pMON13408/ 99 base pair 19 Ala,polypeptide B9 SEQ ID NO: 340 NcoI, EcoRV NcoI, EcoRV 45 Val, SEQ ID NO:272 fragment from 49 Ile pMON13405 Example 78 pMON13423 pMON13408/ 99base pair 19 Ile, polypeptide B10 SEQ ID NO: 341 NcoI, EcoRV NcoI, EcoRV45 Val, SEQ ID NO: 273 fragment from 49 Ile pMON13415 Example 79pMON13424 pMON13409/ 99 base pair 19 Ala, polypeptide B11 SEQ ID NO: 342NcoI, EcoRV NcoI, EcoRV 45 Val, SEQ ID NO: 274 fragment from 49 LeupMON13405 Example 80 pMON13425 pMON13409/ 99 base pair 19 Ile,polypeptide B12 SEQ ID NO: 343 NcoI, EcoRV NcoI, EcoRV 45 Val, SEQ IDNO: 275 fragment from 49 Leu pMON13415 Example 81 pMON13426 pMON13410/99 base pair 19 Ala, polypeptide B13 SEQ ID NO: 344 NcoI, EcoRV NcoI,EcoRV 45 Val, SEQ ID NO: 276 fragment from 49 Asp pMON13405 Example 82pMON13429 pMON13410/ 99 base pair 19 Ile, polypeptide B14 SEQ ID NO: 345NcoI, EcoRV NcoI, EcoRV 45 Val, SEQ ID NO: 277 fragment from 49 AsppMON13415

[1215] TABLE 7 pMON Step one Step one Step two Step two Amino AcidExample number template PCR primer1 PCR primer2 PCR primer1 PCR primer2Substitutions Polypeptide Example 86 pMON13475 pMON13287 18I23A25H42D45V46S50D product from 2341HIN3 42D, 46S, 50D Polypeptide SEQ ID NO:OLIGO# B13 OLIGO# B19 step one OLIGO# B14 # B17 348 SEQ ID NO: 182 SEQID NO: 188 SEQ ID NO: 183 SEQ ID NO: 280 Example 87 pMON13366 pMON132872341NCO 42D45V46S50D product from 2341HIN3 42N, 46S, 50D Polypeptide SEQID NO: OLIGO# B15 OLIGO# B19 step one OLIGO# B14 # B18 349 SEQ ID NO:184 SEQ ID NO: 188 SEQ ID NO: 183 SEQ ID NO: 281 Example 88 pMON13367pMON13287 2341NCO 42A45V46S50D product from 2341HIN3 46S, 50DPolypeptide SEQ ID NO: OLIGO# B15 OLIGO# B17 step one OLIGO# B14 # B19350 SEQ ID NO: 184 SEQ ID NO: 186 SEQ ID NO: 183 SEQ ID NO: 282 Example89 pMON13369 pMON13287 2341NCO 42D45V46S50D product from 2341HIN3 42D,46S, 50D Polypeptide SEQ ID NO: OLIGO# B15 OLIGO# B21 step one OLIGO#B14 # B20 351 SEQ ID NO: 184 SEQ ID NO: 190 SEQ ID NO: 183 SEQ ID NO:283 Example 90 pMON13370 pMON13287 2341NCO 42A45M46S50D product from2341HIN3 45M, 46S, 50D Polypeptide SEQ ID NO: OLIGO# B15 OLIGO# B16 stepone OLIGO# B14 # B21 352 SEQ ID NO: 184 SEQ ID NO: 185 SEQ ID NO: 183SEQ ID NO: 284 Example 91 pMON13373 pMON13287 2341NCO 42D45M46S50Dproduct from 2341HIN3 42D, 45M, 46S Polypeptide SEQ ID NO: OLIGO# B15OLIGO# B18 step one OLIGO# B14 50D # B22 353 SEQ ID NO: 184 SEQ ID NO:187 SEQ ID NO: 183 SEQ ID NO: 285 Example 92 pMON13374 pMON13287 2341NCO42S45M46S50D product from 2341HIN3 42S, 45M, 46S Polypeptide SEQ ID NO:OLIGO# B15 OLIGO# B20 step one OLIGO# B14 50D # B23 354 SEQ ID NO: 184SEQ ID NO: 189 SEQ ID NO: 183 SEQ ID NO: 286

[1216] TABLE 8 re- sulting amino parental acid Example plasmid plasmidoligo pair oligo pair oligo pair oligo pair sub (s). polypeptide Example93 pMON13375 pMON13287/ SO9E16V1 S116VD31 15-119 polypeptide B24 SEQ IDNO: EcoR1, HindIII OLIGO# B50 OLIGO# B52 SEQ ID NO: 287 355 SEQ ID NO:219 SEQ ID NO: 221 SO9E16V3 SECRID33 OLIGO# B51 OLIGO# B53 SEQ ID NO:220 SEQ ID NO: 222 Example 94 pMON13376 pMON13476/ S9E2Q6V1 S116VD3115-119, polypeptide B25 SEQ ID NO: EcoR1,HindIII OLIGO# B54 OLIGO# B5223A, SEQ ID NO: 288 356 SEQ ID NO: 223 SEQ ID NO: 221 112Q S9E2Q6V3SECR1D33 OLIGO# B55 OLIGO# B53 SEQ ID NO: 224 SEQ ID NO: 222 Example 95pMON13377 pMON13475/ S9E2Q6V1 S116VD31 15-119, polypeptide B26 SEQ IDNO: EcoR1, HindIII OLIGO# B54 OLIGO# B52 23A, SEQ ID NO: 289 357 SEQ IDNO: 223 SEQ ID NO: 221 42D, S9E2Q6V3 SECR1D33 46S, OLIGO# B55 OLIGO# B5350D, SEQ ID NO: 224 SEQ ID NO: 222 112Q Example 96 pMON13378 pMON13365/S09E16V1 S116VD31 15-119, polypeptide B27 SEQ ID NO: EcoR1, HindIIIOLIGO# B50 OLIGO# B52 23A SEQ ID NO: 290 358 SEQ ID NO: 219 SEQ ID NO:221 S09E16V3 SECR1D33 OLIGO# B51 OLIGO# B53 SEQ ID NO: 220 SEQ ID NO:222 Example 97 pMON13379 pMON13367/ 9E12Q6V1 120Q123E2 46S, polypeptideB28 SEQ ID NO: EcoR1, HindIII OLIGO# B48 OLIGO# 49 50D, SEQ ID NO: 291359 SEQ ID NO: 217 SED ID NO: 63 112Q 9E12Q6V3 120Q123E4 OLIGO# B49OLIGO# 50 SEQ ID NO: 218 SED ID NO: 64 Example 98 pMON13385 pMON13287/18I25H1 29V32R34S2 42A45V3 29V, polypeptide B29 SEQ ID NO: NcoI, EcoRVOLIGO# 1 OLIGO# B28 OLIGO# 9 32R, SEQ ID NO: 292 360 SEQ ID NO: 15 SEDID NO: 197 SEQ ID NO: 23 34S 18I25H4 29V32R34S5 42A45V6 OLIGO# 2 OLIGO#B29 OLIGO# 10 SEQ ID NO: 16 SED ID NO: 198 SEQ ID NO: 24 Example 99pMON13381 pMON13287/ 73G76A1 82TRP2 87S93S98I3 101A105Q4 82W polypeptideB30 SEQ ID NO: NsiI, EcoRI OLIGO# 41 OLIGO# B44 OLIGO# 35 OLIGO# 43 SEQID NO: 293 361 SEQ ID NO: 55 SEQ ID NO: 213 SEQ ID NO: 49 SEQ ID NO: 5773G76A4 82TRP5 87S93S98I7 101A105Q8 OLIGO# 42 OLIGO# B45 OLIGO# 36OLIGO# 44 SEQ ID NO: 56 SEQ ID NO: 214 SEQ ID NO: 50 SEQ ID NO: 58Example 100 pMON13383 pMON13475/ 9E12Q6V1 120Q123E2 23A, polypeptide B31SEQ ID NO: EcoR1, HindIII OLIGO# B48 OLIGO# 49 42D, SEQ ID NO: 294 362SEQ ID NO: 217 SED ID NO: 63 46S, 9E12Q6V5 120Q123E4 50D OLIGO# B49OLIGO# 50 112Q SEQ ID NO: 218 SED ID NO: 64 Example 101 pMON13384pMON13287/ 9E12Q6V1 120Q123E2 112Q polypeptide B32 SEQ ID NO: EcoR1,HindIII OLIGO# B48 OLIGO# 49 SEQ ID NO: 295 363 SEQ ID NO: 217 SED IDNO: 63 9E12Q6V5 120Q123E4 OLIGO# B49 OLIGO# 50 SEQ ID NO: 218 SED ID NO:64 Example 102 pMON13388 pMON13287/ 50D56S1 60S62V2 67N69E3 50D,polypeptide B33 SEQ ID NO: EcoRV, NsiI OLIGO# B42 OLIGO# 27 OLIGO# 3156S SEQ ID NO: 296 364 SEQ ID NO: 211 SEQ ID NO: 41 SEQ ID NO: 45 50ASP456SER5 67N69E6 OLIGO# B41 OLIGO# B43 OLIGO# 32 SEQ ID NO: 210 SEQ ID NO:212 SEQ ID NO: 46 Example 103 pMON13389 pMON13287/ 18I25H1 29R32A37P242D45MJ 42D, polypeptide B34 SEQ ID NO: NcoI, EcoRV OLIGO# 1 OLIGO# 3OLIGO# B32 45M SEQ ID NO: 297 365 SEQ ID NO: 15 SEQ ID NO: 17 SEQ ID NO:201 18I25H4 29R32A37P5 42D45M6 OLIGO# 2 OLIGO# 4 OLIGO# B33 SEQ ID NO:16 SEQ ID NO: 18 SEQ ID NO: 202 Example 104 pMON13391 pMON13287/ 18I25H134SER1 42A45V3 34S polypeptide B35 SEQ ID NO: NcoI, EcoRV OLIGO# 1OLIGO# B30 OLIGO# 9 SEQ ID NO: 298 366 SEQ ID NO: 15 SEQ ID NO: 199 SEQID NO: 23 18I25H4 34SER5 42A45V6 OLIGO# 2 OLIGO# B31 OLIGO# 10 SEQ IDNO: 16 SEQ ID NO: 200 SEQ ID NO: 24 Example 105 pMON13392 pMON13267/18I25H1 29R32A37P2 42D45V3 42D polypeptide B36 SEQ ID NO: NcoI, EcoRVOLIGO# 1 OLIGO# 3 OLIGO# B34 SEQ ID NO: 299 367 SEQ ID NO: 15 SEQ ID NO:17 SEQ ID NO: 203 18I25H4 29R32A37P5 42D45V6 OLIGO# 2 OLIGO# 4 OLIGO#B35 SEQ ID NO: 16 SEQ ID NO: 18 SEQ ID NO: 204 Example 106 pMON13393PMON13287/ 23ALA1 34SER1 42D45M46S3 23A, polypeptide B37 SEQ ID NO:NcoI, EcoRV OLIGO# B26 OLIGO# B30 OLIGO# B36 34S, SEQ ID NO: 300 368 SEQID NO: 195 SEQ ID NO: 199 SEQ ID NO: 205 42D, 23ALA4 34SER5 42D45M46S645M OLIGO# B27 OLIGO# B31 OLIGO# B37 46S SEQ ID NO: 196 SEQ ID NO: 200SEQ ID NO: 206 Example 107 pMON13394 pMON13287/ 18I25H1 29R32A37P242D45M46S3 42D, polypeptide B38 SEQ ID NO: NcoI, EcoRV OLIGO# 1 OLIGO# 3OLIGO# B36 45M, SEQ ID NO: 301 369 SEQ ID NO: 15 SEQ ID NO: 17 SEQ IDNO: 205 46S 18I25H4 29R32A37P5 42D45M46S6 OLIGO# 2 OLIGO# 4 OLIGO# B37SEQ ID NO: 16 SEQ ID NO: 16 SEQ ID NO: 206 Example 108 pMON13395pMON13267/ 23ALA1 29V32R34S2 42D45V46S3 23A, polypeptide B39 SEQ ID NO:NcoI, EcoRV OLIGO# B26 OLIGO# B28 OLIGO# B38 29V, SEQ ID NO: 302 370 SEQID NO: 195 SED ID NO: 197 SEQ ID NO: 207 32R, 23ALA4 29V32R34S542D45V4656 34S OLIGO# B27 OLIGO# B29 OLIGO# B39 42D, SEQ ID NO: 196 SEDID NO: 198 SEQ ID NO: 208 46S Example 109 pMON13396 pMON13287/ 73G76A179R82Q2 100ARG3 100MET4 100R, polypeptide B40 SEQ ID NO: NsiI, EcoRIOLIGO# 41 OLIGO# 39 SEQ ID NO: OLIGO# B24 101M SEQ ID NO: 303 371 SEQ IDNO: 55 SEQ ID NO: 53 87S93S98I7 SEQ ID NO: 193 73G76A4 79R82Q5 OLIGO# 3610R01M8 OLIGO# 42 OLIGO# 40 SEQ ID NO: 50 OLIGO# B25 SEQ ID NO: 56 SEQID NO: 54 SEQ ID NO: 194 Example 110 pMON13397 pMON13267/ 73G76A1 82TRP2100ARG3 100MET4 82W, polypeptide B41 SEQ ID NO: NsiI, EcoRI OLIGO# 41OLIGO# B44 OLIGO# B22 OLIGO# B24 100R SEQ ID NO: 304 372 SEQ ID NO: 55SEQ ID NO: 213 SEQ ID NO: 191 SEQ ID NO: 193 101M 73G76A4 82TRP587S93S98I7 10R01M8 OLIGO# 42 OLIGO# B45 OLIGO# 36 OLIGO# B25 SEQ ID NO:56 SEQ ID NO: 214 SEQ ID NO: 50 SEQ ID NO: 194 Example 111 pMON13398pMON13287/ 18I25H1 29R32A37P2 42D45V46S3 42D, polypeptide B42 SEQ ID NO:NcoI, EcoRV OLIGO# 1 OLIGO# 3 OLIGO# B38 46S SEQ ID NO: 305 373 SEQ IDNO: 15 SEQ ID NO: 17 SEQ ID NO: 207 18I25H4 29R32A37P5 42D45V46S6 OLIGO#2 OLIGO# 4 OLIGO# B39 SEQ ID NO: 16 SEQ ID NO: 18 SEQ ID NO: 208 Example112 pMON13399 pMON13388/ 23ALA1 29V32R34S2 42D45V46S3 23A, polypeptideB43 SEQ ID NO: NcoI, EcoRV OLIGO# B26 OLIGO# B28 OLIGO# B36 29V SEQ IDNO: 306 374 SEQ ID NO: 195 SED ID NO: 197 SEQ ID NO: 207 32R 23ALA429V32R34S5 42D45V46S6 34S OLIGO# B27 OLIGO# B29 OLIGO# B39 42D SEQ IDNO: 196 SEQ ID NO: 198 SEQ ID NO: 208 46S Example 113 pMON13404pMON13287/ S9E2Q6V1 S116VD31 15-119 polypeptide B44 SEQ ID NO: EcoRI,HindIII OLIGO# B54 OLIGO# B52 112Q SEQ ID NO: 307 375 SEQ ID NO: 223 SEQID NO: 221 S9E2Q6V3 SECR1D33 OLIGO# B55 OLIGO# 53 SEQ ID NO: 224 SEQ IDNO: 222 Example 114 pMON13387 pMON13287/ 50ASP1 60S62V2 67N69E3 50Dpolypeptide B45 SEQ ID NO: EcoRV, NsiI OLIGO# B40 OLIGO# 27 OLIGO# 31SEQ ID NO: 308 376 SEQ ID NO: 209 SEQ ID NO: 41 SEQ ID NO: 45 50ASP460S62V5 67N69E6 OLIGO# B41 OLIGO# 28 OLIGO# 32 SEQ ID NO: 210 SEQ ID NO:42 SEQ ID NO: 46 Example 115 pMON13416 pMON13387/ 18I25H1 29R32A37P242D45V46S3 42D, polypeptide B46 SEQ ID NO: NcoI/EcoRV OLIGO# 1 OLIGO# 3OLIGO# B38 46S SEQ ID NO: 309 377 SEQ ID NO: 15 SEQ ID NO: 17 SEQ ID NO:207 50D 18I25H4 29R32A37P5 42D45V46S6 OLIGO# 2 OLIGO# 4 OLIGO# B39 SEQID NO: 16 SEQ ID NO: 18 SEQ ID NO: 208 Example 116 pMON13417 pMON13387/18I25H1 29R32A37P2 42D45M46S3 42D, polypeptide B47 SEQ ID NO: NcoI/EcoRVOLIGO# 1 OLIGO# 3 OLIGO# B36 45M SEQ ID NO: 310 378 SEQ ID NO: 15 SEQ IDNO: 17 SEQ ID NO: 205 46S 18I25H4 29R32A37P5 42D45M4656 50D OLIGO# 2OLIGO# 4 OLIGO# B37 SEQ ID NO: 16 SEQ ID NO: 18 SEQ ID NO: 206 Example117 pMON13420 pMON13388/ 23ALA1 34SER1 42D45V46S3 23A, polypeptide B48SEQ ID NO: NcoI, EcoRV OLIGO# B26 OLIGO# B30 OLIGO# B38 34S SEQ ID NO:311 379 SEQ ID NO: 195 SEQ ID NO: 199 SEQ ID NO: 207 42D 23ALA4 34SERS42D45V46S6 46S OLIGO# B27 OLIGO# B31 OLIGO# B39 50D SEQ ID NO: 196 SEQID NO: 200 SEQ ID NO: 208 56S Example 118 pMON13421 pMON13388/ 23ALA134SER1 42D45M46S3 23A, polypeptide B49 SEQ ID NO: NcoI, EcoRV OLIGO# B26OLIGO# B30 OLIGO# B36 34S SEQ ID NO: 331 380 SEQ ID NO: 195 SEQ ID NO:199 SEQ ID NO: 205 42D 23ALA4 34SER5 42D45M46S6 45M OLIGO# B27 OLIGO#B31 OLIGO# B37 46S SEQ ID NO: 196 SEQ ID NO: 200 SEQ ID NO: 206 50D 56SExample 119 pMON13432 pMON13387/ 23ALA1 34SER1 42D45M46S3 23A,polypeptide B50 SEQ ID NO: NcoI, EcoRV OLIGO# B26 OLIGO# B30 OLIGO# B3634S SEQ ID NO: 312 381 SEQ ID NO: 195 SEQ ID NO: 199 SEQ ID NO: 205 42D23ALA4 34SER5 42D45M46S6 45M OLIGO# B27 OLIGO# B31 OLIGO# B37 46S SEQ IDNO: 196 SEQ ID NO: 200 SEQ ID NO: 206 50D Example 120 pMON13382pMON13287/ 9E12Q6W1 120Q123E2 112Q, polypeptide B51 SEQ ID NO: EcoRI,HindIII OLIGO# B46 OLIGO# 49 116W SEQ ID NO: 313 382 SEQ ID NO: 215 SEQID NO: 63 9E12Q16W3 120Q123E4 OLIGO# B47 OLIGO# 50 SEQ ID NO: 216 SEQ IDNO: 64

[1217] TABLE 9 Parental Plasmid/ Restriction Amino acid Example No.Plasmid Digest Oligo pair Oligo pair Oligo pair Oligo pair changesPolypeptide Example 124 pMON13400 pMON13288 20P23A1 29I4S7S2 38A5V6S320P 23A Polypeptide C-2 SEQ ID NO: Restriction SEQ ID NO: 232 SEQ ID NO:236 SEQ ID NO: 238 29I 34S SEQ ID NO: 317 384 NcoI-EcoRV 20P23A429I4S7S5 38A5V6S3 37S 38A SEQ ID NO: 233 SEQ ID NO: 237 SEQ ID NO: 23945V 46S Example 125 pMON13402 pMON13288 23L1 29I4S7S2 38A5V6S3 23L 29IPolypeptide C-3 SEQ ID NO: Restriction SEQ ID NO: 234 SEQ ID NO: 236 SEQID NO: 238 34S 37S SEQ ID NO: 318 385 NcoI-EcoRV 23L4 29I4S7S5 38A5V6S338A 45V 46S SEQ ID NO: 235 SEQ ID NO: 237 SEQ ID NO: 239 Example 131pMON13440 pMON13288 18I3A5H1 29I4S7S2 38A5V6S3 18I 23A 25H PolypeptideC-10 SEQ ID NO: Restriction SEQ ID NO: 195 SEQ ID NO: 236 SEQ ID NO: 23829I 34S 37S SEQ ID NO: 319 386 NcoI-EcoRV 18I3A5H4 29I4S7S5 38A5V6S3 38A45V 46S SEQ ID NO: 196 SEQ ID NO: 237 SEQ ID NO: 239 Example 132pMON13451 pMON13288 19I0L3A1 29I4S7S2 38A5V6S3 19I 20L 23A PolypeptideC-11 SEQ ID NO: Restriction SEQ ID NO: 230 SEQ ID NO: 236 SEQ ID NO: 23829I 34S 37S SEQ ID NO: 320 387 NcoI-EcoRV 19I0L3A4 29I4S7S5 38A5V6S3 38A45V 46S SEQ ID NO: 231 SEQ ID NO: 237 SEQ ID NO: 239 Example 130pMON13419 pMON13288 50D51S1 62P3H5S2 67Q3 50D 51S 62P Polypeptide C-8SEQ ID NO: Restriction SEQ ID NO: 240 SEQ ID NO: 244 SEQ ID NO: 248 63HSEQ ID NO: 325 389 EcoRV-NsiI 50D51S4 62P3H5 65S67Q6 65S 67Q SEQ ID NO:241 SEQ ID NO: 246 SEQ ID NO: 247 Example 126 pMON13403 pMON1328850D51S1 62P3H2 67Q3 50D 51S Polypeptide C-4 SEQ ID NO: Restriction SEQID NO: 240 SEQ ID NO: 245 SEQ ID NO: 248 62P 63H SEQ ID NO: 321 388EcoRV-NsiI 50D51S4 62P3H5 67Q6 67Q SEQ ID NO: 241 SEQ ID NO: 246 SEQ IDNO: 249 Example 123 pMON13418 pMON13288 76P1 79S2 5VYWPTT3 101A105Q4 76P79S Polypeptide C-1 SEQ ID NO: Restriction SEQ ID NO: 250 SEQ ID NO: 252SEQ ID NO: 242 SEQ ID NO: 85V 87Y SEQ ID NO: 326 393 Neil-EcoRI 76P579S6 5VYWPTT7 57 88W 91P SEQ ID NO: 251 SEQ ID NO: 253 SEQ ID NO: 243101A105Q8 95T 98T SEQ ID NO: 58 Example 127 pMON13411 pMON13288 09L2Q6S1120Q123E2 109L 112Q Polypeptide C-5 SEQ ID NO: Restriction Seq ID NO:227 SEQ ID NO: 63 116S SEQ ID NO: 322 390 EcoRI- 09L2Q6S3 120Q123E4HindIII SEQ ID NO: 228 SEQ ID NO: 64 Example 128 pMON13412 pMON132889LQS1181 15-118 Polypeptide C-6 SEQ ID NO: Restriction Seq ID NO: 255109L 112Q SEQ ID NO: 323 391 EcoRI- 9LQS1183 116S HindIII SEQ ID NO: 256Example 129 pMON13413 pMON13288 09L2Q6S1 117S2 109L 112Q Polypeptide C-7SEQ ID NO: Restriction Seq ID NO: 227 SEQ ID NO: 229 116S 117S SEQ IDNO: 324 392 EcoRI- 09L2Q6S3 120Q123E4 HindIII SEQ ID NO: 228 SEQ ID NO:64

[1218] TABLE 10 Parental plasmid/ Restriction Restriction Amino AcidExample No Plasmid digest fragment changes Polypeptide Example 133pMON13428 pMON13411 102 bp 76P 79S 85V Polypeptide SEQ ID NsiI-EcoRINsiI-EcoRI 87Y 91P 95T C-9 NO: 394 fragment from 98T 109L SEQ IDpMON13418 112Q 116S NO: 327 Example 134 pMON13459 pMON13428 170 bp 23L29I 34S Polypeptide SEQ ID NcoI-NsiI NcoI-NsiI 37S 38A 45V C-12 NO: 395fragment from 46S 76P 79S SEQ ID pMON13402 85V 87Y 91P NO: 328 95T 98T109L 112Q 116S Example 135 pMON13467 pMON13413 170 bp 23L 29I 34SPolypeptide SEQ ID NcoI-NsiI NcoI-NsiI 37S 38A 45V C-13 NO: 396 fragmentfrom 46S 109L SEQ ID pMON13402 112Q 116S NO: 329 109L 112Q 116S 117SExample 136 pMON13492 pMON13418 170 bp 23L 29I 34S Polypeptide SEQ IDNcoI-NsiI NcoI-NsiI 37S 38A 45V C-14 NO: 397 fragment from 46S 76P 79SSEQ ID pMON13402 85V 87Y 91P NO: 330 95T 98T

What is claimed is:
 1. A human interleukin-3 mutant polypeptide FormulaI: Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn [SEQ IDNO:15]  1           5    10                 15 Cys Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             20   25                 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaAsn Xaa Xaa Xaa Xaa Xaa Xaa              35   40                 45 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             50   55                 60 Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa              65   70                 75 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             80   85                 90 Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa              95  100                105 XaaPhe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa            110  115                120 Xaa Xaa Xaa Gln Gln Thr Thr LeuSer Leu Ala Ile Phe             125  130

wherein Xaa at position 17 is Ser, Lys, Gly, Asp, Met, Gln, or Arg; Xaaat position 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln; Xaa at position19 is Met, Phe, Ile, Arg, Gly, Ala, or Cys; Xaa at position 20 is Ile,Cys, Gln, Glu, Arg, Pro, or Ala; Xaa at position 21 is Asp, Phe, Lys,Arg, Ala, Gly, Glu, Gln, Asn, Thr, Ser or Val; Xaa at position 22 isGlu, Trp, Pro, Ser, Ala, His, Asp, Asn, Gln, Leu, Val or Gly; Xaa atposition 23 is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe, Leu, Ser, or Arg;Xaa at position 24 is Ile, Gly, Val, Arg, Ser, Phe, or Leu; Xaa atposition 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala; Xaa at position 26is His, Thr, Phe, Gly, Arg, Ala, or Trp; Xaa at position 27 is Leu, Gly,Arg, Thr, Ser, or Ala; Xaa at position 28 is Lys, Arg, Leu, Gln, Gly,Pro, Val or Trp; Xaa at position 29 is Gln, Asn, Leu, Pro, Arg, or Val;Xaa at position 30 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys;Xaa at position 31 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln; Xaa atposition 32 is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu; Xaa atposition 33 is Pro, Leu, Gln, Ala, Thr, or Glu; Xaa at position 34 isLeu, Val, Gly, Ser, Lys, Glu, Gln, Thr, Arg, Ala, Phe, Ile or Met; Xaaat position 35 is Leu, Ala, Gly, Asn, Pro, Gln, or Val; Xaa at position36 is Asp, Leu, or Val; Xaa at position 37 is Phe, Ser, Pro, Trp, orIle; Xaa at position 38 is Asn, or Ala; Xaa at position 40 is Leu, Trp,or Arg; Xaa at position 41 is Asn, Cys, Arg, Leu, His, Met, or Pro; Xaaat position 42 is Gly, Asp, Ser, Cys, Asn, Lys, Thr, Leu, Val, Glu, Phe,Tyr, Ile, Met or Ala; Xaa at position 43 is Glu, Asn, Tyr, Leu, Phe,Asp, Ala, Cys, Gln, Arg, Thr, Gly or Ser; Xaa at position 44 is Asp,Ser, Leu, Arg, Lys, Thr, Met, Trp, Glu, Asn, Gln, Ala or Pro; Xaa atposition 45 is Gln, Pro, Phe, Val, Met, Leu, Thr, Lys, Trp, Asp, Asn,Arg, Ser, Ala, Ile, Glu or His; Xaa at position 46 is Asp, Phe, Ser,Thr, Cys, Glu, Asn, Gln, Lys, His, Ala, Tyr, Ile, Val or Gly; Xaa atposition 47 is Ile, Gly, Val, Ser, Arg, Pro, or His; Xaa at position 48is Leu, Ser, Cys, Arg, Ile, His, Phe, Glu, Lys, Thr, Ala, Met, Val orAsn; Xaa at position 49 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp;Xaa at position 50 is Glu, Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala, Ile,Val, His, Phe, Met or Gln; Xaa at position 51 is Asn, Arg, Met, Pro,Ser, Thr, or His; Xaa at position 52 is Asn, His, Arg, Leu, Gly, Ser, orThr; Xaa at position 53 is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, orMet; Xaa at position 54 is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn, Lys,His, Ala or Leu; Xaa at position 55 is Arg, Thr, Val, Ser, Leu, or Gly;Xaa at position 56 is Pro, Gly, Cys, Ser, Gln, Glu, Arg, His, Thr, Ala,Tyr, Phe, Leu, Val or Lys; Xaa at position 57 is Asn or Gly; Xaa atposition 58 is Leu, Ser, Asp, Arg, Gln, Val, or Cys; Xaa at position 59is Glu Tyr, His, Leu, Pro, or Arg; Xaa at position 60 is Ala, Ser, Pro,Tyr, Asn, or Thr; Xaa at position 61 is Phe, Asn, Glu, Pro, Lys, Arg, orSer; Xaa at position 62 is Asn His, Val, Arg, Pro, Thr, Asp, or Ile; Xaaat position 63 is Arg, Tyr, Trp, Lys, Ser, His, Pro, or Val; Xaa atposition 64 is Ala, Asn, Pro, Ser, or Lys; Xaa at position 65 is Val,Thr, Pro, His, Leu, Phe, or Ser; Xaa at position 66 is Lys, Ile, Arg,Val, Asn, Glu, or Ser; Xaa at position 67 is Ser, Ala, Phe, Val, Gly,Asn, Ile, Pro, or His; Xaa at position 68 is Leu, Val, Trp, Ser, Ile,Phe, Thr, or His; Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg,Trp, Gly, or Leu; Xaa at position 70 is Asn, Leu, Val, Trp, Pro, or Ala;Xaa at position 71 is Ala, Met, Leu, Pro, Arg, Glu, Thr, Gln, Trp, orAsn; Xaa at position 72 is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp;Xaa at position 73 is Ala, Glu, Asp, Leu, Ser, Gly, Thr, or Arg; Xaa atposition 74 is Ile, Met, Thr, Pro, Arg, Gly, Ala; Xaa at position 75 isGlu, Lys, Gly, Asp, Pro, Trp, Arg, Ser, Gln, or Leu; Xaa at position 76is Ser, Val, Ala, Asn, Trp, Glu, Pro, Gly, or Asp; Xaa at position 77 isIle, Ser, Arg, Thr, or Leu; Xaa at position 78 is Leu, Ala, Ser, Glu,Phe, Gly, or Arg; Xaa at position 79 is Lys, Thr, Asn, Met, Arg, Ile,Gly, or Asp; Xaa at position 80 is Asn, Trp, Val, Gly, Thr, Leu, Glu, orArg; Xaa at position 81 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys;Xaa at position 82 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn, His, Thr,Ser, Ala, Tyr, Phe, Ile, Met or Val; Xaa at position 83 is Pro, Ala,Thr, Trp, Arg, or Met; Xaa at position 84 is Cys, Glu, Gly, Arg, Met, orVal; Xaa at position 85 is Leu, Asn, Val, or Gln; Xaa at position 86 isPro, Cys, Arg, Ala, or Lys; Xaa at position 87 is Leu, Ser, Trp, or Gly;Xaa at position 88 is Ala, Lys, Arg, Val, or Trp; Xaa at position 89 isThr, Asp, Cys, Leu, Val, Glu, His, Asn, or Ser; Xaa at position 90 isAla, Pro, Ser, Thr, Gly, Asp, Ile, or Met; Xaa at position 91 is Ala,Pro, Ser, Thr, Phe, Leu, Asp, or His; Xaa at position 92 is Pro, Phe,Arg, Ser, Lys, His, Ala, Gly, Ile or Leu; Xaa at position 93 is Thr,Asp, Ser, Asn, Pro, Ala, Leu, or Arg; Xaa at position 94 is Arg, Ile,Ser, Glu, Leu, Val, Gln, Lys, His, Ala, or Pro; Xaa at position 95 isHis, Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn, Lys, Ser, Ala, Trp, Phe,Ile, or Tyr; Xaa at position 96 is Pro, Lys, Tyr, Gly, Ile, or Thr; Xaaat position 97 is Ile, Val, Lys, Ala, or Asn; Xaa at position 98 is His,Ile, Asn, Leu, Asp, Ala, Thr, Glu, Gln, Ser, Phe, Met, Val, Lys, Arg,Tyr or Pro; Xaa at position 99 is Ile, Leu, Arg, Asp, Val, Pro, Gln,Gly, Ser, Phe, or His; Xaa at position 100 is Lys, Tyr, Leu, His, Arg,Ile, Ser, Gln, or Pro; Xaa at position 101 is Asp, Pro, Met, Lys, His,Thr, Val, Tyr, Glu, Asn, Ser, Ala, Gly, Ile, Leu, or Gln; Xaa atposition 102 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro; Xaa at position103 is Asp, or Ser; Xaa at position 104 is Trp, Val, Cys, Tyr, Thr, Met,Pro, Leu, Gln, Lys, Ala, Phe, or Gly; Xaa at position 105 is Asn, Pro,Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His; Xaa atposition 106 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro; Xaa atposition 108 is Arg, Lys, Asp, Leu, Thr, Ile, Gln, His, Ser, Ala or Pro;Xaa at position 109 is Arg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly; Xaa atposition 110 is Lys, Ala, Asn, Thr, Leu, Arg, Gln, His, Glu, Ser, Ala,or Trp; Xaa at position 111 is Leu, Ile, Arg, Asp, or Met; Xaa atposition 112 is Thr, Val, Gln, Tyr, Glu, His, Ser, or Phe; Xaa atposition 113 is Phe, Ser, Cys, His, Gly, Trp, Tyr, Asp, Lys, Leu, Ile,Val or Asn; Xaa at position 114 is Tyr, Cys, His, Ser, Trp, Arg, or Leu;Xaa at position 115 is Leu, Asn, Val, Pro, Arg, Ala, His, Thr, Trp, orMet; Xaa at position 116 is Lys, Leu, Pro, Thr, Met, Asp, Val, Glu, Arg,Trp, Ser, Asn, His, Ala, Tyr, Phe, Gln, or Ile; Xaa at position 117 isThr, Ser, Asn, Ile, Trp, Lys, or Pro; Xaa at position 118 is Leu, Ser,Pro, Ala, Glu, Cys, Asp, or Tyr; Xaa at position 119 is Glu, Ser, Lys,Pro, Leu, Thr, Tyr, or Arg; Xaa at position 120 is Asn, Ala, Pro, Leu,His, Val, or Gln; Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys,Asp, or Gly; Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro,His, Ile, Tyr, or Cys; Xaa at position 123 is Ala, Met, Glu, His, Ser,Pro, Tyr, or Leu; and which can additionally have Met- preceding theamino acid in position 1; and wherein from 1 to 14 amino acids can bedeleted from the N-terminus and/or from 1 to 15 amino acids can bedeleted from the C-terminus; and wherein from 4 to 44 of the amino acidsdesignated by Xaa are different from the corresponding amino acids ofnative (1-133) human interleukin-3.
 2. A human interleukin-3 mutantpolypeptide of the Formula II: Ala Pro Met Thr Gln Thr Thr Ser Leu LysThr Ser Trp Val Asn [SEQ ID NO:16] 1           5   10                  15 Cys Xaa Xaa Xaa Xaa Xaa Glu XaaXaa Xaa Xaa Leu Xaa Xaa Xaa             20    25                 30 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa Xaa Glu Xaa Xaa            35    40                 45 Xaa Xaa Leu Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Asn Leu Xaa Xaa             50    55                 60 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa            65    70                 75 Xaa Xaa Leu Xaa Xaa Xaa Xaa XaaCys Xaa Pro Xaa Xaa Xaa Xaa             80    85                 90 XaaXaa Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asp Xaa Xaa            95   100                105 Xaa Phe Xaa Xaa Lys Leu Xaa PheXaa Xaa Xaa Xaa Leu Xaa Xaa             110  115                120 XaaXaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe             125  130

wherein Xaa at position 17 is Ser, Gly, Asp, Met, or Gln; Xaa atposition 18 is Asn, His, Leu, Ile, Phe, Arg, or Gln; Xaa at position 19is Met, Phe, Ile, Arg, or Ala; Xaa at position 20 is Ile or Pro; Xaa atposition 21 is Asp or Glu; Xaa at position 23 is Ile, Val, Ala, Leu, orGly; Xaa at position 24 is Ile, Val, Phe, or Leu; Xaa at position 25 isThr, His, Gly, Gln, Arg, Pro, or Ala; Xaa at position 26 is His, Phe,Gly, Arg, or Ala; Xaa at position 28 is Lys, Leu, Gln, Gly, Pro, or Val;Xaa at position 29 is Gln, Asn, Leu, Arg, or Val; Xaa at position 30 isPro, His, Thr, Gly, or Gln; Xaa at position 31 is Pro, Asp, Gly, Ala,Arg, Leu, or Gln; Xaa at position 32 is Leu, Arg, Gln, Asn, Gly, Ala, orGlu; Xaa at position 33 is Pro, Leu, Gln, Ala, or Glu; Xaa at position34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met;Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val; Xaa at position36 is Asp or Leu; Xaa at position 37 is Phe, Ser, Pro, Trp, or Ile; Xaaat position 38 is Asn or Ala; Xaa at position 41 is Asn, Cys, Arg, His,Met, or Pro; Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile,Leu, Met, Tyr, Val or Arg; Xaa at position 44 is Asp or Glu; Xaa atposition 45 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn, Glu, Ser, or Trp;Xaa at position 46 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu, His,Ile, Lys, Tyr, Val or Gly; Xaa at position 47 is Ile, Val, or His; Xaaat position 49 is Met, Asn, or Asp; Xaa at position 50 is Glu, Thr, Ala,Asn, Ser or Asp; Xaa at position 51 is Asn, Arg, Met, Pro, Ser, Thr, orHis; Xaa at position 52 is Asn or Gly; Xaa at position 53 is Leu, Met,or Phe; Xaa at position 54 is Arg, Ala, or Ser; Xaa at position 55 isArg, Thr, Val, Leu, or Gly; Xaa at position 56 is Pro, Gly, Cys, Ser,Gln, Ala, Arg, Asn, Glu, His, Leu, Thr, Val or Lys; Xaa at position 59is Glu, Tyr, His, Leu, or Arg; Xaa at position 60 is Ala, Ser, Asn, orThr; Xaa at position 61 is Phe or Ser; Xaa at position 62 is Asn, Val,Pro, Thr, or Ile; Xaa at position 63 is Arg, Tyr, Lys, Ser, His, or Val;Xaa at position 64 is Ala or Asn; Xaa at position 65 is Val, Thr, Leu,or Ser; Xaa at position 66 is Lys, Ile, Arg, Val, Asn, Glu, or Ser; Xaaat position 67 is Ser, Phe, Val, Gly, Asn, Ile, or His; Xaa at position68 is Leu, Val, Ile, Phe, or His; Xaa at position 69 is Gln, Ala, Pro,Thr, Glu, Arg, or Gly; Xaa at position 70 is Asn or Pro; Xaa at position71 is Ala, Met, Pro, Arg, Glu, Thr, or Gln; Xaa at position 72 is Ser,Glu, Met, Ala, His, Asn, Arg, or Asp; Xaa at position 73 is Ala, Glu,Asp, Leu, Ser, Gly, Thr, Arg, or Pro; Xaa at position 74 is Ile or Met;Xaa at position 75 is Glu, Gly, Asp, Ser, or Gln; Xaa at position 76 isSer, Val, Ala, Asn, Glu, Pro, Gly, or Asp; Xaa at position 77 is Ile,Ser, or Leu; Xaa at position 79 is Lys, Thr, Gly, Asn, Met, Arg, Ile,Gly, or Asp; Xaa at position 80 is Asn, Val, Gly, Thr, Leu, Glu, or Arg;Xaa at position 81 is Leu, or Val; Xaa at position 82 is Leu, Gln, Trp,Arg, Asp, Ala, Asn, Glu, His, Met, Phe, Ser, Thr, Tyr or Val; Xaa atposition 83 is Pro, Ala, Thr, Trp, or Met; Xaa at position 85 is Leu orVal; Xaa at position 87 is Leu or Ser; Xaa at position 88 is Ala, Arg,or Trp; Xaa at position 89 is Thr, Asp, Glu, His, Asn, or Ser; Xaa atposition 90 is Ala, Asp, or Met; Xaa at position 91 is Ala, Pro, Ser,Thr, Phe, Leu, or Asp; Xaa at position 92 is Pro or Ser; Xaa at position93 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg; Xaa at position 95 is His,Pro, Arg, Val, Leu, Gly, Asn, Ile, Phe, Ser or Thr; Xaa at position 96is Pro or Tyr; Xaa at position 97 is Ile, Val, or Ala; Xaa at position98 is His, Ile, Asn, Leu, Asp, Ala, Thr, Leu, Arg, Gln, Glu, Lys, Met,Ser, Tyr, Val or Pro; Xaa at position 99 is Ile, Leu, Val, or Phe; Xaaat position 100 is Lys, Leu, His, Arg, Ile, Gln, Pro, or Ser; Xaa atposition 101 is Asp, Pro, Met, Lys, His, Thr, Val, Asn, Ile, Leu or Tyr;Xaa at position 102 is Gly, Glu, Lys, or Ser; Xaa at position 104 isTrp, Val, Tyr, Met, or Leu; Xaa at position 105 is Asn, Pro, Ala, Phe,Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His; Xaa at position 106 isGlu, Ser, Ala, or Gly; Xaa at position 108 is Arg, Ala, Gln, Ser or Lys;Xaa at position 109 is Arg, Thr, Glu, Leu, Ser, or Gly; Xaa at position112 is Thr, Val, Gln, Glu, His, or Ser; Xaa at position 114 is Tyr orTrp; Xaa at position 115 is Leu or Ala; Xaa at position 116 is Lys, Thr,Met, Val, Trp, Ser, Leu, Ala, Asn, Gln, His, Met, Phe, Tyr or Ile; Xaaat position 117 is Thr, Ser, or Asn; Xaa at position 119 is Glu, Ser,Pro, Leu, Thr, or Tyr; Xaa at position 120 is Asn, Pro, Leu, His, Val,or Gln; Xaa at position 121 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, orGly; Xaa at position 122 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile,Tyr, or Cys; Xaa at position 123 is Ala, Met, Glu, His, Ser, Pro, Tyr,or Leu; and which can additionally have Met- preceding the amino acid inposition 1; and wherein from 1 to 14 amino acids can be deleted from theN-terminus and/or from 1 to 15 amino acids can be deleted from theC-terminus; and wherein from 4 to 44 of the amino acids designated byXaa are different from the corresponding amino acids of native (1-133)human interleukin-3.
 3. A human interleukin-3 mutant polypeptideaccording to claim 2 of the Formula III: Ala Pro Met Thr Gln Thr Thr SerLeu Lys Thr Ser Trp Val Asn [SEQ ID NO:17] 1           5    10                 15 Cys Xaa Xaa Xaa Ile Xaa Glu XaaXaa Xaa Xaa Leu Lys Xaa Xaa              20   25                 30 XaaXaa Xaa Xaa Xaa Asp Xaa Xaa Asn Leu Asn Xaa Glu Xaa Xaa             35   40                 45 Xaa Ile Leu Met Xaa Xaa Asn LeuXaa Xaa Xaa Asn Leu Glu Xaa              50   55                 60 PheXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile Glu             65   70                 75 Xaa Xaa Leu Xaa Xaa Leu Xaa XaaCys Xaa Pro Xaa Xaa Thr Ala              80   85                 90 XaaPro Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa Xaa             95  100                105 Xaa Phe Xaa Xaa Lys Leu Xaa PheXaa Xaa Xaa Xaa Leu Glu Xaa             110  115                120 XaaXaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe             125  130

wherein Xaa at position 17 is Ser, Gly, Asp, Met, or Gln; Xaa atposition 18 is Asn, His, or Ile; Xaa at position 19 is Met or Ile; Xaaat position 21 is Asp or Glu; Xaa at position 23 is Ile, Ala, Leu, orGly; Xaa at position 24 is Ile, Val, or Leu; Xaa at position 25 is Thr,His, Gln, or Ala; Xaa at position 26 is His or Ala; Xaa at position 29is Gln, Asn, or Val; Xaa at position 30 is Pro, Gly, or Gln; Xaa atposition 31 is Pro, Asp, Gly, or Gln; Xaa at position 32 is Leu, Arg,Gln, Asn, Gly, Ala, or Glu; Xaa at position 33 is Pro or Glu; Xaa atposition 34 is Leu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe,Thr or Met; Xaa at position 35 is Leu, Ala, Asn, Pro, Gln, or Val; Xaaat position 37 is Phe, Ser, Pro, or Trp; Xaa at position 38 is Asn orAla; Xaa at position 42 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met,Tyr or Arg; Xaa at position 44 is Asp or Glu; Xaa at position 45 is Gln,Val, Met, Leu, Thr, Ala, Asn, Glu, Ser or Lys; Xaa at position 46 isAsp, Phe, Ser, Thr, Ala, Asn Gln, Glu, His, Ile, Lys, Tyr, Val or Cys;Xaa at position 50 is Glu, Ala, Asn, Ser or Asp; Xaa at position 51 isAsn, Arg, Met, Pro, Ser, Thr, or His; Xaa at position 54 is Arg or Ala;Xaa at position 55 is Arg, Thr, Val, Leu, or Gly; Xaa at position 56 isPro, Gly, Ser, Gln, Ala, Arg, Asn, Glu, Leu, Thr, Val or Lys; Xaa atposition 60 is Ala or Ser; Xaa at position 62 is Asn, Pro, Thr, or Ile;Xaa at position 63 is Arg or Lys; Xaa at position 64 is Ala or Asn; Xaaat position 65 is Val or Thr; Xaa at position 66 is Lys or Arg; Xaa atposition 67 is Ser, Phe, or His; Xaa at position 68 is Leu, Ile, Phe, orHis; Xaa at position 69 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly; Xaa atposition 71 is Ala, Pro, or Arg; Xaa at position 72 is Ser, Glu, Arg, orAsp; Xaa at position 73 is Ala or Leu; Xaa at position 76 is Ser, Val,Ala, Asn, Glu, Pro, or Gly; Xaa at position 77 is Ile or Leu; Xaa atposition 79 is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, or Asp; Xaa atposition 80 is Asn, Gly, Glu, or Arg; Xaa at position 82 is Leu, Gln,Trp, Arg, Asp, Ala, Asn, Glu, His, Ile, Met, Phe, Ser, Thr, Tyr or Val;Xaa at position 83 is Pro or Thr; Xaa at position 85 is Leu or Val; Xaaat position 87 is Leu or Ser; Xaa at position 88 is Ala or Trp; Xaa atposition 91 is Ala or Pro; Xaa at position 93 is Thr, Asp, Ser, Pro,Ala, Leu, or Arg; Xaa at position 95 is His, Pro, Arg, Val, Leu, Gly,Asn, Phe, Ser or Thr; Xaa at position 96 is Pro or Tyr; Xaa at position97 is Ile or Val; Xaa at position 98 is His, Ile, Asn, Leu, Ala, Thr,Leu, Arg, Gln, Leu, Lys, Met, Ser, Tyr, Val or Pro; Xaa at position 99is Ile, Leu, or Val; Xaa at position 100 is Lys, Arg, Ile, Gln, Pro, orSer; Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Pro, Asn, Ile,Leu or Tyr; Xaa at position 104 is Trp or Leu; Xaa at position 105 isAsn, Pro, Ala, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His; Xaa atposition 106 is Glu or Gly; Xaa at position 108 is Arg, Ala, or Ser; Xaaat position 109 is Arg, Thr, Glu, Leu, or Ser; Xaa at position 112 isThr, Val, or Gln; Xaa at position 114 is Tyr or Trp; Xaa at position 115is Leu or Ala; Xaa at position 116 is Lys, Thr, Val, Trp, Ser, Ala, His,Met, Phe, Tyr or Ile; Xaa at position 117 is Thr or Ser; Xaa at position120 is Asn, Pro, Leu, His, Val, or Gln; Xaa at position 121 is Ala, Ser,Ile, Asn, Pro, Asp, or Gly; Xaa at position 122 is Gln, Ser, Met, Trp,Arg, Phe, Pro, His, Ile, Tyr, or Cys; Xaa at position 123 is Ala, Met,Glu, His, Ser, Pro, Tyr, or Leu; and which can additionally have Met-preceding the amino acid in position 1; and wherein from 1 to 14 aminoacids can be deleted from the N-terminus and/or from 1 to 15 amino acidscan be deleted from the C-terminus; and wherein from 4 to 35 of theamino acids designated by Xaa are different from the corresponding aminoacids of native (1-133) human interleukin-3.
 4. A human interieukin-3mutant polypeptide according to claim 3 of the Formula IV: Ala Pro MetThr Gln Thr Thr Ser Leu Lys Thr Ser Trp Val Asn [SEQ ID NO:18] 1           5    10                 15 Cys Xaa Xaa Met Ile Asp Glu XaaIle Xaa Xaa Leu Lys Xaa Xaa             20    25                 30 ProXaa Pro Xaa Xaa Asp Phe Xaa Asn Leu Asn Xaa Glu Asp Xaa           35     40                 45 Xaa Ile Leu Met Xaa Xaa Asn LeuArg Xaa Xaa Asn Leu Glu Ala            50     55                 60 PheXaa Arg Xaa Xaa Lys Xaa Xaa Xaa Asn Ala Ser Ala Ile Glu           65    70                  75 Xaa Xaa Leu Xaa Xaa Leu Xaa ProCys Leu Pro Xaa Xaa Thr Ala            80     85                 90 XaaPro Xaa Arg Xaa Pro Ile Xaa Xaa Xaa Xaa Gly Asp Trp Xaa            95   100                105 Glu Phe Xaa Xaa Lys Leu Xaa PheTyr Leu Xaa Xaa Leu Glu Xaa             110  115                120 XaaXaa Xaa Gln Gln Thr Thr Leu Ser Leu Ala Ile Phe             125  130

wherein Xaa at position 17 is Ser, Gly, Asp, or Gln; Xaa at position 18is Asn, His, or Ile; Xaa at position 23 is Ile, Ala, Leu, or Gly; Xaa atposition 25 is Thr, His, or Gln; Xaa at position 26 is His or Ala; Xaaat position 29 is Gln or Asn; Xaa at position 30 is Pro or Gly; Xaa atposition 32 is Leu, Arg, Asn, or Ala; Xaa at position 34 is Leu, Val,Ser, Ala, Arg, Gln, Glu, Ile, Phe, Thr, or Met; Xaa at position 35 isLeu, Ala, Asn, or Pro; Xaa at position 38 is Asn or Ala; Xaa at position42 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyr or Arg; Xaa atposition 45 is Gln, Val, Met, Leu, Ala, Asn, Glu, or Lys; Xaa atposition 46 is Asp, Phe, Ser, Gln, Glu, His, Val or Thr; Xaa at position50 is Glu Asn, Ser or Asp; Xaa at position 51 is Asn, Arg, Pro, Thr, orHis; Xaa at position 55 is Arg, Leu, or Gly; Xaa at position 56 is Pro,Gly, Ser, Ala, Asn, Val, Leu or Gln; Xaa at position 62 is Asn, Pro, orThr; Xaa at position 64 is Ala or Asn; Xaa at position 65 is Val or Thr;Xaa at position 67 is Ser or Phe; Xaa at position 68 is Leu or Phe; Xaaat position 69 is Gln, Ala, Glu, or Arg; Xaa at position 76 is Ser, Val,Asn, Pro, or Gly; Xaa at position 77 is Ile or Leu; Xaa at position 79is Lys, Gly, Asn, Met, Arg, Ile, or Gly; Xaa at position 80 is Asn, Gly,Glu, or Arg; Xaa at position 82 is Leu, Gln, Trp, Arg, Asp, Asn, Glu,His, Met, Phe, Ser, Thr, Tyr or Val; Xaa at position 87 is Leu or Ser;Xaa at position 88 is Ala or Trp; Xaa at position 91 is Ala or Pro; Xaaat position 93 is Thr, Asp, or Ala; Xaa at position 95 is His, Pro, Arg,Val, Gly, Asn, Ser or Thr; Xaa at position 98 is His, Ile, Asn, Ala,Thr, Gln, Glu, Lys, Met, Ser, Tyr, Val or Leu; Xaa at position 99 is Ileor Leu; Xaa at position 100 is Lys or Arg; Xaa at position 101 is Asp,Pro, Met, Lys, Thr, His, Pro, Asn, Ile, Leu or Tyr; Xaa at position 105is Asn, Pro, Ser, Ile or Asp; Xaa at position 108 is Arg, Ala, or Ser;Xaa at position 109 is Arg, Thr, Glu, Leu, or Ser; Xaa at position 112is Thr or Gln; Xaa at position 116 is Lys, Val, Trp, Ala, His, Phe, Tyror Ile; Xaa at position 117 is Thr or Ser; Xaa at position 120 is Asn,Pro, Leu, His, Val, or Gln; Xaa at position 121 is Ala, Ser, Ile, Pro,or Asp; Xaa at position 122 is Gln, Met, Trp, Phe, Pro, His, Ile, orTyr; Xaa at position 123 is Ala, Met, Glu, Ser, or Leu; and which canadditionally have Met- preceding the amino acid in position 1; andwherein from 1 to 14 amino acids can be deleted from the N-terminusand/or from 1 to 15 amino acids can be deleted from the C-terminus; andwherein from 4 to 44 of the amino acids designated by Xaa are differentfrom the corresponding amino acids of native (1-133) humaninterleukin-3.
 5. The human interleukin-3 mutant polypeptide of claim 1wherein 1-15 amino acids are deleted from the C-terminus and/or 1-14amino acids are deleted from the N-terminus.
 6. The human interleukin-3mutant polypeptide of claim 1 wherein; Xaa at position 42 is Gly, Asp,Ser, Ile, Leu, Met, Tyr, or Ala; Xaa at position 45 is Gln, Val, Met orAsn; Xaa at position 46 is Asp, Ser, Gln, His or Val; Xaa at position 50is Glu or Asp; Xaa at position 51 is Asn, Pro or Thr; Xaa at position 62is Asn or Pro; Xaa at position 76 is Ser, or Pro; Xaa at position 82 isLeu, Trp, Asp, Asn Glu, His, Phe, Ser or Tyr; Xaa at position 95 is His,Arg, Thr, Asn or Ser; Xaa at position 98 is His, Ile, Leu, Ala, Gln,Lys, Met, Ser, Tyr or Val; Xaa at position 100 is Lys or Arg; Xaa atposition 101 is Asp, Pro, His, Asn, Ile or Leu; Xaa at position 105 isAsn, or Pro; Xaa at position 108 is Arg, Ala, or Ser; Xaa at position116 is Lys, Val, Trp, Ala, His, Phe, or Tyr; Xaa at position 121 is Ala,or Ile; Xaa at position 122 is Gln, or Ile; and Xaa at position 123 isAla, Met or Glu.
 7. A (15-125) human interleukin-3 mutant polypeptide ofthe Formula V: Asn Cys Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa [SEQ ID NO:19]  1           5    10                 15 Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Xaa Xaa             20   25                 30 Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa              35   40                 45 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             50   55                 60 Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa              65   70                 75 XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa             80   85                 90 Xaa Xaa Phe Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa              95  100                105 XaaXaa Xaa Xaa Gln Gln             110

wherein Xaa at position 3 is Ser, Lys, Gly, Asp, Met, Gln, or Arg; Xaaat position 4 is Asn, His, Leu, Ile, Phe, Arg, or Gln; Xaa at position 5is Met, Phe, Ile, Arg, Gly, Ala, or Cys; Xaa at position 6 is Ile, Cys,Gln, Glu, Arg, Pro, or Ala; Xaa at position 7 is Asp, Phe, Lys, Arg,Ala, Gly, Glu, Gln, Asn, Thr, Ser or Val; Xaa at position 8 is Glu, Trp,Pro, Ser, Ala, His, Asp, Asn, Gln, Leu, Val, or Gly; Xaa at position 9is Ile, Val, Ala, Leu, Gly, Trp, Lys, Phe, Leu, Ser, or Arg; Xaa atposition 10 is Ile, Gly, Val, Arg, Ser, Phe, or Leu; Xaa at position 11is Thr, His, Gly, Gln, Arg, Pro, or Ala; Xaa at position 12 is His, Thr,Phe, Gly, Arg, Ala, or Trp; Xaa at position 13 is Leu, Gly, Arg, Thr,Ser, or Ala; Xaa at position 14 is Lys, Arg, Leu, Gln, Gly, Pro, Val orTrp; Xaa at position 15 is Gln, Asn, Leu, Pro, Arg, or Val; Xaa atposition 16 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, or Lys; Xaa atposition 17 is Pro, Asp, Gly, Ala, Arg, Leu, or Gln; Xaa at position 18is Leu, Val, Arg, Gln, Asn, Gly, Ala, or Glu; Xaa at position 19 is Pro,Leu, Gln, Ala, Thr, or Glu; Xaa at position 20 is Leu, Val, Gly, Ser,Lys, Glu, Gln, Thr, Arg, Ala, Phe, Ile or Met; Xaa at position 21 isLeu, Ala, Gly, Asn, Pro, Gln, or Val; Xaa at position 22 is Asp, Leu, orVal; Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile; Xaa at position24 is Asn, or Ala; Xaa at position 26 is Leu, Trp, or Arg; Xaa atposition 27 is Asn, Cys, Arg, Leu, His, Met, Pro; Xaa at position 28 isGly, Asp, Ser, Cys, Ala, Lys, Asn, Thr, Leu, Val, Glu, Phe, Tyr, Ile orMet; Xaa at position 29 is Glu, Asn, Tyr, Leu, Phe, Asp, Ala, Cys, Gln,Arg, Thr, Gly or Ser; Xaa at position 30 is Asp, Ser, Leu, Arg, Lys,Thr,Met, Trp, Glu, Asn, Gln, Ala or Pro; Xaa at position 31 is Gln, Pro,Phe, Val, Met, Leu, Thr, Lys, Asp, Asn, Arg, Ser, Ala, Ile, Glu, His orTrp; Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Glu, Asn, Gln, Lys,His, Ala, Tyr, Ile, Val or Gly; Xaa at position 33 is Ile, Gly, Val,Ser, Arg, Pro, or His; Xaa at position 34 is Leu, Ser, Cys, Arg, Ile,His, Phe, Glu, Lys, Thr, Ala, Met, Val or Asn; Xaa at position 35 isMet, Arg, Ala, Gly, Pro, Asn, His, or Asp; Xaa at position 36 is Glu,Leu, Thr, Asp, Tyr, Lys, Asn, Ser, Ala, Ile, Val, His, Phe, Met or Gln;Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, or His; Xaa atposition 38 is Asn, His, Arg, Leu, Gly, Ser, or Thr; Xaa at position 39is Leu, Thr, Ala, Gly, Glu, Pro, Lys, Ser, Met, or; Xaa at position 40is Arg, Asp, Ile, Ser, Val, Thr, Gln, Asn, Lys, His, Ala or Leu; Xaa atposition 41 is Arg, Thr, Val, Ser, Leu, or Gly; Xaa at position 42 isPro, Gly, Cys, Ser, Gln, Glu, Arg, His, Thr, Ala, Tyr, Phe, Leu, Val orLys; Xaa at position 43 is Asn or Gly; Xaa at position 44 is Leu, Ser,Asp, Arg, Gln, Val, or Cys; Xaa at position 45 is Glu Tyr, His, Leu,Pro, or Arg; Xaa at position 46 is Ala, Ser, Pro, Tyr, Asn, or Thr; Xaaat position 47 is Phe, Asn, Glu, Pro, Lys, Arg, or Ser; Xaa at position48 is Asn, His, Val, Arg, Pro, Thr, Asp, or Ile; Xaa at position 49 isArg, Tyr, Trp, Lys, Ser, His, Pro, or Val; Xaa at position 50 is Ala,Asn, Pro, Ser, or Lys; Xaa at position 51 is Val, Thr, Pro, His, Leu,Phe, or Ser; Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser;Xaa at position 53 is Ser, Ala, Phe, Val, Gly, Asn, Ile, Pro, or His;Xaa at position 54 is Leu, Val, Trp, Ser, Ile, Phe, Thr, or His; Xaa atposition 55 is Gln, Ala, Pro, Thr, Glu, Arg, Trp, Gly, or Leu; Xaa atposition 56 is Asn, Leu, Val, Trp, Pro, or Ala; Xaa at position 57 isAla, Met, Leu, Pro, Arg, Glu, Thr, Gln, Trp, or Asn; Xaa at position 58is Ser, Glu, Met, Ala, His, Asn, Arg, or Asp; Xaa at position 59 is Ala,Glu, Asp, Leu, Ser, Gly, Thr, or Arg; Xaa at position 60 is Ile, Met,Thr, Pro, Arg, Gly, Ala; Xaa at position 61 is Glu, Lys, Gly, Asp, Pro,Trp, Arg, Ser, Gln, or Leu; Xaa at position 62 is Ser, Val, Ala, Asn,Trp, Glu, Pro, Gly, or Asp; Xaa at position 63 is Ile, Ser, Arg, Thr, orLeu; Xaa at position 64 is Leu, Ala, Ser, Glu, Phe, Gly, or Arg; Xaa atposition 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, or Asp; Xaa atposition 66 is Asn, Trp, Val, Gly, Thr, Leu, Glu, or Arg; Xaa atposition 67 is Leu, Gln, Gly, Ala, Trp, Arg, Val, or Lys; Xaa atposition 68 is Leu, Gln, Lys, Trp, Arg, Asp, Glu, Asn, His, Thr, Ser,Ala, Tyr, Phe, Ile, Met or Val; Xaa at position 69 is Pro, Ala, Thr,Trp, Arg, or Met; Xaa at position 70 is Cys, Glu, Gly, Arg, Met, or Val;Xaa at position 71 is Leu, Asn, Val, or Gln; Xaa at position 72 is Pro,Cys, Arg, Ala, or Lys; Xaa at position 73 is Leu, Ser, Trp, or Gly; Xaaat position 74 is Ala, Lys, Arg, Val, or Trp; Xaa at position 75 is Thr,Asp, Cys, Leu, Val, Glu, His, Asn, or Ser; Xaa at position 76 is Ala,Pro, Ser, Thr, Gly, Asp, Ile, or Met; Xaa at position 77 is Ala, Pro,Ser, Thr, Phe, Leu, Asp, or His; Xaa at position 78 is Pro, Phe, Arg,Ser, Lys, His, Ala, Gly, Ile or Leu; Xaa at position 79 is Thr, Asp,Ser, Asn, Pro, Ala, Leu, or Arg; Xaa at position 80 is Arg, Ile, Ser,Glu, Leu, Val, Gln, Lys, His, Ala or Pro; Xaa at position 81 is His,Gln, Pro, Arg, Val, Leu, Gly, Thr, Asn, Lys, Ser, Ala, Trp, Phe, Ile orTyr; Xaa at position 82 is Pro, Lys, Tyr, Gly, Ile, or Thr; Xaa atposition 83 is Ile, Val, Lys, Ala, or Asn; Xaa at position 84 is His,Ile, Asn, Leu, Asp, Ala, Thr, Glu, Gln, Ser, Phe, Met, Val, Lys, Arg,Tyr or Pro; Xaa at position 85 is Ile, Leu, Arg, Asp, Val, Pro, Gln,Gly, Ser, Phe, or His; Xaa at position 86 is Lys, Tyr, Leu, His, Arg,Ile, Ser, Gln, Pro; Xaa at position 87 is Asp, Pro, Met, Lys, His, Thr,Val, Tyr, Glu, Asn, Ser, Ala, Gly, Ile, Leu or Gln; Xaa at position 88is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro; Xaa at position 89 is Asp, orSer; Xaa at position 90 is Trp, Val, Cys, Tyr, Thr, Met, Pro, Leu, Gln,Lys, Ala, Phe, or Gly; Xaa at position 91 is Asn, Pro, Ala, Phe, Ser,Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His; Xaa at position 92 is Glu,Ser, Ala, Lys, Thr, Ile, Gly, or Pro; Xaa at position 94 is Arg, Lys,Asp, Leu, Thr, Ile, Gln, His, Ser, Ala, or Pro; Xaa at position 95 isArg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly; Xaa at position 96 is Lys,Asn, Thr, Leu, Gln, Arg, His, Glu, Ser, Ala or Trp; Xaa at position 97is Leu, Ile, Arg, Asp, or Met; Xaa at position 98 is Thr, Val, Gln, Tyr,Glu, His, Ser, or Phe; Xaa at position 99 is Phe, Ser, Cys, His, Gly,Trp, Tyr, Asp, Lys, Leu, Ile, Val or Asn; Xaa at position 100 is Tyr,Cys, His, Ser, Trp, Arg, or Leu; Xaa at position 101 is Leu, Asn, Val,Pro, Arg, Ala, His, Thr, Trp, or Met; Xaa at position 102 is Lys, Leu,Pro, Thr, Met, Asp, Val, Glu, Arg, Trp, Ser, Asn, His, Ala, Tyr, Phe,Gln, or Ile; Xaa at position 103 is Thr, Ser, Asn, Ile, Trp, Lys, orPro; Xaa at position 104 is Leu, Ser, Pro, Ala, Glu, Cys, Asp, or Tyr;Xaa at position 105 is Glu, Ser, Lys, Pro, Leu, Thr, Tyr, or Arg; Xaa atposition 106 is Asn, Ala, Pro, Leu, His, Val, or Gln; Xaa at position107 is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly; Xaa at position 108 isGln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys; Xaa atposition 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu; and which canadditionally have Met- or Met-Ala- preceding the amino acid in position1; and wherein from 4 to 44 of the amino acids designated by Xaa aredifferent from the corresponding native amino acids of (1-133) humaninterleukin-3; or a polypeptide having substantially the same structureand substantially the same biological activity.
 8. A (15-125) humaninterleukin-3 mutant polypeptide of the Formula VI: Asn Cys Xaa Xaa XaaXaa Xaa Glu Xaa Xaa Xaa Xaa Leu Xaa Xaa [SEQ ID NO:20] 1           5   10                  15 Xaa Xaa Xaa Xaa Xaa Xaa Xaa XaaXaa Asn Leu Xaa Xaa Glu Xaa              20  25                  30 XaaXaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Leu Xaa             35  40                  45 Xaa Xaa Xea Xaa Xaa Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Xaa Xaa              50  55                  60 XaaXaa Xaa Leu Xaa Xaa Xaa Xaa Xaa Cys Xaa Pro Xaa Xaa Xaa             65  70                  75 Xaa Xaa Xaa Xaa Arg Xaa Xaa XaaXaa Xaa Xaa Xaa Xaa Asp Xaa              80  85                  90 XaaXaa Phe Xaa Xaa Lys Leu Xaa Phe Xaa Xaa Xaa Xaa Leu Xaa             95 100                 105 Xaa Xaa Xaa Xaa Gln Gln            110

wherein Xaa at position 3 is Ser, Gly, Asp, Met, or Gln; Xaa at position4 is Asn, His, Leu, Ile, Phe, Arg, or Gln; Xaa at position 5 is Met,Phe, Ile, Arg, or Ala; Xaa at position 6 is Ile or Pro; Xaa at position7 is Asp, or Glu; Xaa at position 9 is Ile, Val, Ala, Leu, or Gly; Xaaat position 10 is Ile, Val, Phe, or Leu; Xaa at position 11 is Thr, His,Gly, Gln, Arg, Pro, or Ala; Xaa at position 12 is His, Phe, Gly, Arg, orAla; Xaa at position 14 is Lys, Leu, Gln, Gly, Pro, or Val; Xaa atposition 15 is Gln, Asn, Leu, Arg, or Val; Xaa at position 16 is Pro,His, Thr, Gly, or Gln; Xaa at position 17 is Pro, Asp, Gly, Ala, Arg,Leu, or Gln; Xaa at position 18 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu;Xaa at position 19 is Pro, Leu, Gln, Ala, or Glu; Xaa at position 20 isLeu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met; Xaaat position 21 is Leu, Ala, Asn, Pro, Gln, or Val; Xaa at position 22 isAsp or Leu; Xaa at position 23 is Phe, Ser, Pro, Trp, or Ile; Xaa atposition 24 is Asn or Ala; Xaa at position 27 is Asn, Cys, Arg, His,Met, or Pro; Xaa at position 28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile,Leu, Met, Tyr, or Arg; Xaa at position 30 is Asp, or Glu; Xaa atposition 31 is Gln, Val, Met, Leu, Thr, Lys, Ala, Asn Glu, Ser or Trp;Xaa at position 32 is Asp, Phe, Ser, Thr, Cys, Ala, Asn, Gln, Glu, His,Ile, Lys, Tyr, Val or Gly; Xaa at position 33 is Ile, Val, or His; Xaaat position 35 is Met, Asn, or Asp; Xaa at position 36 is Glu, Thr, Ala,Asn, Ser or Asp; Xaa at position 37 is Asn, Arg, Met, Pro, Ser, Thr, orHis; Xaa at position 38 is Asn or Gly; Xaa at position 39 is Leu, Met,or Phe; Xaa at position 40 is Arg, Ala or Ser; Xaa at position 41 isArg, Thr, Val, Leu, or Gly; Xaa at position 42 is Pro, Gly, Cys, Ser,Gln, Ala, Arg, Asn, Glu, His, Leu, Thr, Val or Lys; Xaa at position 45is Glu, Tyr, His, Leu, or Arg; Xaa at position 46 is Ala, Ser, Asn, orThr; Xaa at position 47 is Phe or Ser; Xaa at position 48 is Asn, Val,Pro, Thr, or Ile; Xaa at position 49 is Arg, Tyr, Lys, Ser, His, or Val;Xaa at position 50 is Ala or Asn; Xaa at position 51 is Val, Thr, Leu,or Ser; Xaa at position 52 is Lys, Ile, Arg, Val, Asn, Glu, or Ser; Xaaat position 53 is Ser, Phe, Val, Gly, Asn, Ile, or His; Xaa at position54 is Leu, Val, Ile, Phe, or His; Xaa at position 55 is Gln, Ala, Pro,Thr, Glu, Arg, or Gly; Xaa at position 56 is Asn or Pro; Xaa at position57 is Ala, Met, Pro, Arg, Glu, Thr, or Gln; Xaa at position 58 is Ser,Glu, Met, Ala, His, Asn, Arg, or Asp; Xaa at position 59 is Ala, Glu,Asp, Leu, Ser, Gly, Thr, Arg, or Pro; Xaa at position 60 is Ile or Met;Xaa at position 61 is Glu, Gly, Asp, Ser, or Gln; Xaa at position 62 isSer, Val, Ala, Asn, Glu, Pro, Gly, or Asp; Xaa at position 63 is Ile,Ser, or Leu; Xaa at position 65 is Lys, Thr, Gly, Asn, Met, Arg, Ile, orAsp; Xaa at position 66 is Asn, Val, Gly, Thr, Leu, Glu, or Arg; Xaa atposition 67 is Leu, or Val; Xaa at position 68 is Leu, Gln, Trp, Arg,Asp, Ala, Asn, Glu, His, Met, Phe, Ser, Thr, Tyr or Val; Xaa at position69 is Pro, Ala, Thr, Trp, or Met; Xaa at position 71 is Leu or Val; Xaaat position 73 is Leu or Ser; Xaa at position 74 is Ala, Arg, or Trp;Xaa at position 75 is Thr, Asp, Glu, His, Asn, or Ser; Xaa at position76 is Ala, Asp, or Met; Xaa at position 77 is Ala, Pro, Ser, Thr, Phe,Leu, or Asp; Xaa at position 78 is Pro or Ser; Xaa at position 79 isThr, Asp, Ser, Pro, Ala, Leu, or Arg; Xaa at position 81 is His, Pro,Arg, Val, Leu, Gly, Asn, Ile, Phe, Ser or Thr; Xaa at position 82 is Proor Tyr; Xaa at position 83 is Ile, Val, or Ala; Xaa at position 84 isHis, Ile, Asn, Leu, Asp, Ala, Thr, Arg, Gln, Glu, Lys, Met, Ser, Tyr,Val or Pro; Xaa at position 85 is Ile, Leu, Val, or Phe; Xaa at position86 is Lys, Leu, His, Arg, Ile, Gln, Pro or Ser; Xaa at position 87 isAsp, Pro, Met, Lys, His, Thr, Val, Asn, Ile, Leu or Tyr; Xaa at position88 is Gly, Glu, Lys, or Ser; Xaa at position 90 is Trp, Val, Tyr, Met,or Leu; Xaa at position 91 is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr,Leu, Lys, Ile, Asp, or His; Xaa at position 92 is Glu, Ser, Ala, or Gly;Xaa at position 94 is Arg, Ala, Gln, Ser or Lys; Xaa at position 95 isArg, Thr, Glu, Leu, Ser, or Gly; Xaa at position 98 is Thr, Val, Gln,Glu, His, or Ser; Xaa at position 100 is Tyr or Trp; Xaa at position 101is Leu or Ala; Xaa at position 102 is Lys, Thr, Met, Val, Trp, Ser, Leu,Ala, Asn, Gln, His, Met, Phe, Tyr or Ile; Xaa at position 103 is Thr,Ser, or Asn; Xaa at position 105 is Glu, Ser, Pro, Leu, Thr, or Tyr; Xaaat position 106 is Asn, Pro, Leu, His, Val, or Gln; Xaa at position 107is Ala, Ser, Ile, Asn, Pro, Lys, Asp, or Gly; Xaa at position 108 isGln, Ser, Met, Trp, Arg, Phe, Pro, His, Ile, Tyr, or Cys; Xaa atposition 109 is Ala, Met, Glu, His, Ser, Pro, Tyr, or Leu; and which canadditionally have Met- or Met-Ala- preceding the amino acid in position1; and wherein from 4 to 44 of the amino acids designated by Xaa aredifferent from the corresponding amino acids of native (1-133) humaninterleukin-3; or a polypeptide having substantially the same structureand substantially the same biological activity.
 9. A (15-125) humaninterleukin-3 mutant polypeptide according to claim 7 of the FormulaVII: Asn Cys Xaa Xaa Xaa Ile Xaa Gln Xaa Xaa Xaa Xaa Leu Lys Xaa [SEQ IDNO:21]  1           5   10                  15 Xaa Xaa Xaa Xaa Xaa XaaAsp Xaa Xaa Asn Leu Asn Xaa Glu Xaa             20  25                  30 Xaa Xaa Ile Leu Met Xaa Xaa AsnLeu Xaa Xaa Xaa Asn Leu Gln              35  40                  45 XaaPhe Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Asn Xaa Xaa Xaa Ile             50  55                  60 Gln Xaa Xaa Leu Xaa Xaa Leu XaaXaa Cys Xaa Pro Xaa Xaa Thr              65  70                  75 AlaXaa Pro Xaa Arg Xaa Xaa Xaa Xaa Xaa Xaa Xaa Gly Asp Xaa             80  85                  90 Xaa Xaa Phe Xaa Xaa Lys Leu XaaPhe Xaa Xaa Xaa Xaa Leu Glu              95 100                 105 XaaXaa Xaa Xaa Gln Gln             110

wherein Xaa at position 3 is Ser, Gly, Asp, Met, or Gln; Xaa at position4 is Asn, His, or Ile; Xaa at position 5 is Met or Ile; Xaa at position7 is Asp or Glu; Xaa at position 9 is Ile, Ala, Leu, or Gly; Xaa atposition 10 is Ile, Val, or Leu; Xaa at position 11 is Thr, His, Gln, orAla; Xaa at position 12 is His or Ala; Xaa at position 15 is Gln, Asn,or Val; Xaa at position 16 is Pro, Gly, or Gln; Xaa at position 17 isPro, Asp, Gly, or Gln; Xaa at position 18 is Leu, Arg, Gln, Asn, Gly,Ala, or Glu; Xaa at position 19 is Pro or Glu; Xaa at position 20 isLeu, Val, Gly, Ser, Lys, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met; Xaaat position 21 is Leu, Ala, Asn, Pro, Gln, or Val; Xaa at position 23 isPhe, Ser, Pro, or Trp; Xaa at position 24 is Asn or Ala; Xaa at position28 is Gly, Asp, Ser, Cys, Ala, Asn, Ile, Leu, Met Tyr or Arg; Xaa atposition 30 is Asp or Glu; Xaa at position 31 is Gln, Val, Met, Leu,Thr, Ala, Asn, Glu, Ser or Lys; Xaa at position 32 is Asp, Phe, Ser,Thr, Ala, Asn, Gln, Glu, His, Ile, Lys, Tyr, Val or Cys; Xaa at position36 is Glu, Ala, Asn, Ser or Asp; Xaa at position 37 is Asn, Arg, Met,Pro, Ser, Thr, or His; Xaa at position 40 is Arg or Ala; Xaa at position41 is Arg, Thr, Val, Leu, or Gly; Xaa at position 42 is Pro, Gly, Ser,Gln, Ala, Arg, Asn, Glu, Leu, Thr, Val or Lys; Xaa at position 46 is Alaor Ser; Xaa at position 48 is Asn, Pro, Thr, or Ile; Xaa at position 49is Arg or Lys; Xaa at position 50 is Ala or Asn; Xaa at position 51 isVal or Thr; Xaa at position 52 is Lys or Arg; Xaa at position 53 is Ser,Phe, or His; Xaa at position 54 is Leu, Ile, Phe, or His; Xaa atposition 55 is Gln, Ala, Pro, Thr, Glu, Arg, or Gly; Xaa at position 57is Ala, Pro, or Arg; Xaa at position 58 is Ser, Glu, Arg, or Asp; Xaa atposition 59 is Ala or Leu; Xaa at position 62 is Ser, Val, Ala, Asn,Glu, Pro, or Gly; Xaa at position 63 is Ile or Leu; Xaa at position 65is Lys, Thr, Gly, Asn, Met, Arg, Ile, Gly, or Asp; Xaa at position 66 isAsn, Gly, Glu, or Arg; Xaa at position 68 is Leu, Gln, Trp, Arg, Asp,Ala, Asn, Glu, His, Ile, Met, Phe, Ser, Thr, Tyr or Val; Xaa at position69 is Pro or Thr; Xaa at position 71 is Leu or Val; Xaa at position 73is Leu or Ser; Xaa at position 74 is Ala or Trp; Xaa at position 77 isAla or Pro; Xaa at position 79 is Thr, Asp, Ser, Pro, Ala, Leu, or Arg;Xaa at position 81 is His, Pro, Arg, Val, Leu, Gly, Asn, Phe, Ser orThr; Xaa at position 82 is Pro or Tyr; Xaa at position 83 is Ile or Val;Xaa at position 84 is His, Ile, Asn, Leu, Ala, Thr, Leu, Arg, Gln, Leu,Lys, Met, Ser, Tyr, Val or Pro; Xaa at position 85 is Ile, Leu, or Val;Xaa at position 86 is Lys, Arg, Ile, Gln, Pro, or Ser; Xaa at position87 is Asp, Pro, Met, Lys, His, Thr, Asn, Ile, Leu or Tyr; Xaa atposition 90 is Trp or Leu; Xaa at position 91 is Asn, Pro, Ala, Ser,Trp, Gln, Tyr, Leu, Lys, Ile, Asp, or His; Xaa at position 92 is Glu, orGly; Xaa at position 94 is Arg, Ala, or Ser; Xaa at position 95 is Arg,Thr, Glu, Leu, or Ser; Xaa at position 98 is Thr, Val, or Gln; Xaa atposition 100 is Tyr or Trp; Xaa at position 101 is Leu or Ala; Xaa atposition 102 is Lys, Thr, Val, Trp, Ser, Ala, His, Met, Phe, Tyr or Ile;Xaa at position 103 is Thr or Ser; Xaa at position 106 is Asn, Pro, Leu,His, Val, or Gln; Xaa at position 107 is Ala, Ser, Ile, Asn, Pro, Asp,or Gly; Xaa at position 108 is Gln, Ser, Met, Trp, Arg, Phe, Pro, His,Ile, Tyr, or Cys; Xaa at position 109 is Ala, Met, Glu, His, Ser, Pro,Tyr, or Leu; which can additionally have Met- or Met-Ala- preceding theamino acid in position 1; and wherein from 4 to 35 of the amino acidsdesignated by Xaa are different from the corresponding amino acids ofnative human interleukin-3.
 10. A (15-125) human interieukin-3 mutantpolypeptide according to claim 7 of the Formula VIII: Asn Cys Xaa XaaMet Ile Asp Glu Xaa Ile Xaa Xaa Leu Lys Xaa [SEQ ID NO:22] 1           5   10                  15 Xaa Pro Xaa Pro Xaa Xaa Asp PheXaa Asn Leu Asn Xaa Gln Asp              20  25                  30 XaaXaa Ile Leu Met Xaa Xaa Asn Leu Arg Xaa Xaa Asn Leu Gln             35  40                  45 Ala Phe Xaa Arg Xaa Xaa Lys XaaXaa Xaa Asn Ala Ser Ala Ile              50  55                  60 GluXaa Xaa Leu Xaa Xaa Leu Xaa Pro Cys Leu Pro Xaa Xaa Thr             65  70                  75 Ala Xaa Pro Xaa Arg Xaa Pro IleXaa Xaa Xaa Xaa Gly Asp Trp              80  85                  90 XaaGln Phe Xaa Xaa Lys Leu Xaa Phe Tyr Leu Xaa Xaa Leu Gln              95100                 105 Xaa Xaa Xaa Xaa Gln Gln             110

wherein Xaa at position 3 is Ser, Gly, Asp, or Gln; Xaa at position 4 isAsn, His, or Ile; Xaa at position 9 is Ile, Ala, Leu, or Gly; Xaa atposition 11 is Thr, His, or Gln; Xaa at position 12 is His or Ala; Xaaat position 15 is Gln or Asn; Xaa at position 16 is Pro or Gly; Xaa atposition 18 is Leu, Arg, Asn, or Ala; Xaa at position 20 is Leu, Val,Ser, Ala, Arg, Gln, Glu, Ile, Phe, Thr or Met; Xaa at position 21 isLeu, Ala, Asn, or Pro; Xaa at position 24 is Asn or Ala; Xaa at position28 is Gly, Asp, Ser, Ala, Asn, Ile, Leu, Met, Tyr or Arg; Xaa atposition 31 is Gln, Val, Met, Leu, Ala, Asn, Glu or Lys; Xaa at position32 is Asp, Phe, Ser, Ala, Gln, Glu, His, Val or Thr; Xaa at position 36is Gln, Asn, Ser or Asp; Xaa at position 37 is Asn, Arg, Pro, Thr, orHis; Xaa at position 41 is Arg, Leu, or Gly; Xaa at position 42 is Pro,Gly, Ser, Ala, Asn, Val, Leu or Gln; Xaa at position 48 is Asn, Pro, orThr; Xaa at position 50 is Ala or Asn; Xaa at position 51 is Val or Thr;Xaa at position 53 is Ser or Phe; Xaa at position 54 is Leu or Phe; Xaaat position 55 is Gln, Ala, Glu, or Arg; Xaa at position 62 is Ser, Val,Asn, Pro, or Gly; Xaa at position 63 is Ile or Leu; Xaa at position 65is Lys, Asn, Met, Arg, Ile, or Gly; Xaa at position 66 is Asn, Gly, Glu,or Arg; Xaa at position 68 is Leu, Gln, Trp, Arg, Asp, Asn, Glu, His,Met, Phe, Ser, Thr, Tyr or Val; Xaa at position 73 is Leu or Ser; Xaa atposition 74 is Ala or Trp; Xaa at position 77 is Ala or Pro; Xaa atposition 79 is Thr, Asp, or Ala; Xaa at position 81 is His, Pro, Arg,Val, Gly, Asn, Ser or Thr; Xaa at position 84 is His, Ile, Asn, Ala,Thr, Arg, Gln, Glu, Lys, Met, Ser, Tyr, Val or Leu; Xaa at position 85is Ile or Leu; Xaa at position 86 is Lys or Arg; Xaa at position 87 isAsp, Pro, Met, Lys, His, Pro, Asn, Ile, Leu or Tyr; Xaa at position 91is Asn, Pro, Ser, Ile or Asp; Xaa at position 94 is Arg, Ala, or Ser;Xaa at position 95 is Arg, Thr, Glu, Leu, or Ser; Xaa at position 98 isThr or Gln; Xaa at position 102 is Lys, Val, Trp, or Ile; Xaa atposition 103 is Thr, Ala, His, Phe, Tyr or Ser; Xaa at position 106 isAsn, Pro, Leu, His, Val, or Gln; Xaa at position 107 is Ala, Ser, Ile,Pro, or Asp; Xaa at position 108 is Gln, Met, Trp, Phe, Pro, His, Ile,or Tyr; Xaa at position 109 is Ala, Met, Glu, Ser, or Leu; and which canadditionally have Met- or Met-Ala- preceding the amino acid in position1; and wherein from 4 to 26 of the amino acids designated by Xaa aredifferent from the corresponding amino acids of native (1-133) humaninterleukin-3; or a polypeptide having substantially the same structureand substantially the same biological activity.
 11. A (15-125) humaninterleukin-3 mutant polypeptide of claim 7 wherein: Xaa at position 17is Ser, Lys, Asp, Met, Gln, or Arg; Xaa at position 18 is Asn, His, Leu,Ile, Phe, Arg, or Gln; Xaa at position 19 is Met, Arg, Gly, Ala, or Cys;Xaa at position 20 is Ile, Cys, Gln, Glu, Arg, Pro, or Ala; Xaa atposition 21 is Asp, Phe, Lys, Arg, Ala, Gly, or Val; Xaa at position 22is Glu, Trp, Pro, Ser, Ala, His, or Gly; Xaa at position 23 is Ile, Ala,Gly, Trp, Lys, Leu, Ser, or Arg; Xaa at position 24 is Ile, Gly, Arg, orSer; Xaa at position 25 is Thr, His, Gly, Gln, Arg, Pro, or Ala; Xaa atposition 26 is His, Thr, Phe, Gly, Ala, or Trp; Xaa at position 27 isLeu, Gly, Arg, Thr, Ser, or Ala; Xaa at position 28 is Lys, Leu, Gln,Gly, Pro, Val or Trp; Xaa at position 29 is Gln, Asn, Loh, Pro, Arg, orVal; Xaa at position 30 is Pro, His, Thr, Gly, Asp, Gln, Ser, Leu, orLys; Xaa at position 31 is Pro, Asp, Gly, Arg, Leu, or Gln; Xaa atposition 32 is Leu, Arg, Gln, Asn, Gly, Ala, or Glu; Xaa at position 33is Pro, Leu, Gln, Thr, or Glu; Xaa at position 34 is Leu, Gly, Ser, orLys; Xaa at position 35 is Leu, Ala, Gly, Asn, Pro, or Gln; Xaa atposition 36 is Asp, Leu, or Val; Xaa at position 37 is Phe, Ser, or Pro;Xaa at position 38 is Asn, or Ala; Xaa at position 40 is Leu, Trp, orArg; Xaa at position 41 is Asn, Cys, Arg, Leu, His, Met, Pro; Xaa atposition 42 is Gly, Asp, Ser, Cys, or Ala; Xaa at position 42 is Glu,Asn, Tyr, Leu, Phe, Asp, Ala, Cys, or Ser; Xaa at position 44 is Asp,Ser, Leu, Arg, Lys, Thr, Met, Trp, or Pro; Xaa at position 45 is Gln,Pro, Phe, Val, Met, Leu, Thr, Lys, or Trp; Xaa at position 46 is Asp,Phe, Ser, Thr, Cys, or Gly; Xaa at position 47 is Ile, Gly, Ser, Arg,Pro, or His; Xaa at position 48 is Leu, Ser, Cys, Arg, His, Phe, or Asn;Xaa at position 49 is Met, Arg, Ala, Gly, Pro, Asn, His, or Asp; Xaa atposition 50 is Glu, Leu, Thr, Asp, or Tyr; Xaa at position 51 is Asn,Arg, Met, Pro, Ser, Thr, or His; Xaa at position 52 is Asn, His, Arg,Leu, Gly, Ser, or Thr; Xaa at position 53 is Leu, Thr, Ala, Gly, Glu,Pro, Lys, Ser, or; Xaa at position 54 is Arg, Asp, Ile, Ser, Val, Thr,Gln, or Leu; Xaa at position 55 is Arg, Thr, Val, Ser, Leu, or Gly; Xaaat position 56 is Pro, Gly, Cys, Ser, Gln, or Lys; Xaa at position 57 isAsn or Gly; Xaa at position 58 is Leu, Ser, Asp, Arg, Gln, Val, or Cys;Xaa at position 59 is Glu Tyr, His, Leu, Pro, or Arg; Xaa at position 60is Ala, Ser, Tyr, Asn, or Thr; Xaa at position 61 is Phe, Asn, Gln, Pro,Lys, Arg, or Ser; Xaa at position 62 is Asn His, Val, Arg, Pro, Thr, orIle; Xaa at position 63 is Arg, Tyr, Trp, Ser, Pro, or Val; Xaa atposition 64 is Ala, Asn, Ser, or Lys; Xaa at position 65 is Val, Thr,Pro, His, Leu, Phe, or Ser; Xaa at position 66 is Lys, Ile, Val, Asn,Gln, or Ser; Xaa at position 67 is Ser, Ala, Phe, Val, Gly, Asn, Ile,Pro, or His; Xaa at position 68 is Leu, Val, Trp, Ser, Thr, or His; Xaaat position 69 is Gln, Ala, Pro, Thr, Arg, Trp, Gly, or Leu; Xaa atposition 70 is Asn, Leu, Val, Trp, Pro, or Ala; Xaa at position 71 isAla, Met, Leu, Arg, Glu, Thr, Gln, Trp, or Asn; Xaa at position 72 isSer, Glu, Met, Ala, His, Asn, Arg, or Asp; Xaa at position 73 is Ala,Gln, Asp, Leu, Ser, Gly, Thr, or Arg; Xaa at position 74 is Ile, Thr,Pro, Arg, Gly, Ala; Xaa at position 75 is Glu, Lys, Gly, Asp, Pro, Trp,Arg, Ser, or Leu; Xaa at position 76 is Ser, Val, Ala, Asn, Trp, Gln,Pro, Gly, or Asp; Xaa at position 77 is Ile, Ser, Arg, or Thr; Xaa atposition 78 is Leu, Ala, Ser, Glu, Gly, or Arg; Xaa at position 79 isLys, Thr, Gly, Asn, Met, Ile, or Asp; Xaa at position 80 is Asn, Trp,Val, Gly, Thr, Leu, or Arg; Xaa at position 81 is Leu, Gln, Gly, Ala,Trp, Arg, or Lys; Xaa at position 82 is Leu, Gln, Lys, Trp, Arg, or Asp;Xaa at position 83 is Pro, Thr, Trp, Arg, or Met; Xaa at position 84 isCys, Gln, Gly, Arg, Met, or Val; Xaa at position 85 is Leu, Asn, or Gln;Xaa at position 86 is Pro, Cys, Arg, Ala, or Lys; Xaa at position 87 isLeu, Ser, Trp, or Gly; Xaa at position 88 is Ala, Lys, Arg, Val, or Trp;Xaa at position 89 is Thr, Asp, Cys, Leu, Val, Glu, His, or Asn; Xaa atposition 90 is Ala, Ser, Asp, Ile, or Met; Xaa at position 91 is Ala,Ser, Thr, Phe, Leu, Asp, or His; Xaa at position 92 is Pro, Phe, Arg,Ser, Lys, His, or Leu; Xaa at position 93 is Thr, Asp, Ser, Asn, Pro,Ala, Leu, or Arg; Xaa at position 94 is Arg, Ile, Ser, Glu, Leu, Val, orPro; Xaa at position 95 is His, Gln, Pro, Val, Leu, Thr or Tyr; Xaa atposition 96 is Pro, Lys, Tyr, Gly, Ile, or Thr; Xaa at position 97 isIle, Lys, Ala, or Asn; Xaa at position 98 is His, Ile, Asn, Leu, Asp,Ala, Thr, or Pro; Xaa at position 99 is Ile, Arg, Asp, Pro, Gln, Gly,Phe, or His; Xaa at position 100 is Lys, Tyr, Leu, His, Ile, Ser, Gln,or Pro; Xaa at position 101 is Asp, Pro, Met, Lys, His, Thr, Val, Tyr,or Gln; Xaa at position 102 is Gly, Leu, Glu, Lys, Ser, Tyr, or Pro; Xaaat position 103 is Asp, or Ser; Xaa at position 104 is Trp, Val, Cys,Tyr, Thr, Met, Pro, Leu, Gln, Lys, Ala, Phe, or Gly; Xaa at position 105is Asn, Pro, Ala, Phe, Ser, Trp, Gln, Tyr, Leu, Lys, Ile, or His; Xaa atposition 106 is Glu, Ser, Ala, Lys, Thr, Ile, Gly, or Pro; Xaa atposition 108 is Arg, Asp, Leu, Thr, Ile, or Pro; Xaa at position 109 isArg, Thr, Pro, Glu, Tyr, Leu, Ser, or Gly.
 12. The human interleukin-3mutant polypetide of claim 7: wherein; Xaa at position 28 is Gly, Asp,Ser, Ile, Leu, Met, Tyr, or Ala; Xaa at position 31 is Gln, Val, Met orAsn; Xaa at position 32 is Asp, Ser, Ala, Gln, His or Val; Xaa atposition 36 is Glu or Asp; Xaa at position 37 is Asn, Pro or Thr; Xaa atposition 48 is Asn or Pro; Xaa at position 62 is Ser, or Pro; Xaa atposition 68 is Leu, Trp, Asp, Asn Glu, His, Phe, Ser or Tyr; Xaa atposition 81 is His, Arg, Thr, Asn or Ser; Xaa at position 84 is His,Ile, Leu, Ala, Arg, Gln, Lys, Met, Ser, Tyr or Val; Xaa at position 86is Lys or Arg; Xaa at position 87 is Asp, Pro, His, Asn, Ile or Leu; Xaaat position 91 is Asn, or Pro; Xaa at position 94 is Arg, Ala, or Ser;Xaa at position 102 is Lys, Val, Trp, Ala, His, Phe, or Tyr; Xaa atposition 107 is Ala, or Ile; Xaa at position 108 is Gln, or Ile; and Xaaat position 109 is Ala, Met or Glu.
 13. A polypeptide of the formula       1               5                  10 (Met)_(m)-Ala Pro Met ThrGln Thr Thr Ser Leu Lys Thr [SEQ ID NO:29]            15                 20 Ser Trp Val Asn Cys Ser Xaa Xaa XaaAsp Gln Ile Ile 25                  30                 35 Xaa His LeuLys Xaa Pro Pro Xaa Pro Xaa Leu Asp Xaa        40                  45             50 Xaa Asn Leu Asn Xaa GlnAsp Xaa Asp Ile Leu Xaa Glu                 55                 60 XaaAsn Leu Arg Xaa Xaa Asn Leu Xaa Xaa Phe Xaa Xaa    65                  70              75 Ala Xaa Lys Xaa Leu Xaa AsnAla Ser Xaa Ile Gln Xaa             80                  85 Ile Leu XaaAsn Leu Xaa Pro Cys Xaa Pro Xaa Xaa Thr90                  95                 100 Ala Xaa Pro Xaa Arg Xaa ProIle Xaa Ile Xaa Xaa Gly        105                110                 115 Asp Trp Xaa Glu PheArg Xaa Lys Leu Xaa Phe Tyr Leu                 120                125Xaa Xaa Leu Glu Xaa Ala Gln Xaa Gln Gln Thr Thr Leu     130 Ser Leu AlaIle Phe

wherein m is 0 or 1; Xaa at position 18 is Asn or Ile; Xaa at position19 is Met, Ala or Ile; Xaa at position 20 is Ile, Pro or Ile; Xaa atposition 23 is Ile, Ala or Leu; Xaa at position 25 is Thr or His; Xaa atposition 29 is Gln, Arg, Val or Ile; Xaa at position 32 is Leu, Ala, Asnor Arg; Xaa at position 34 is Leu or Ser; Xaa at position 37 is Phe,Pro, or Ser; Xaa at position 38 is Asn or Ala; Xaa at position 42 isGly, Ala, Ser, Asp or Asn; Xaa at position 45 is Gln, Val, or Met; Xaaat position 46 is Asp or Ser; Xaa at position 49 is Met, Ile, Leu orAsp; Xaa at position 50 is Glu or Asp; Xaa at position 51 is Asn Arg orSer; Xaa at position 55 is Arg, Leu, or Thr; Xaa at position 56 is Proor Ser; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala orSer; Xaa at position 62 is Asn, Val or Pro; Xaa at position 63 is Arg orHis; Xaa at position 65 is Val or Ser; Xaa at position 67 is Ser, Asn,His or Gln; Xaa at position 69 is Gln or Glu; Xaa at position 73 is Alaor Gly; Xaa at position 76 is Ser, Ala or Pro; Xaa at position 79 isLys, Arg or Ser; Xaa at position 82 is Leu, Glu, Val or Trp; Xaa atposition 85 is Leu or Val; Xaa at position 87 is Leu, Ser, Tyr; Xaa atposition 88 is Ala or Trp; Xaa at position 91 is Ala or Pro; Xaa atposition 93 is Pro or Ser; Xaa at position 95 is His or Thr; Xaa atposition 98 is His, Ile, or Thr; Xaa at position 100 is Lys or Arg; Xaaat position 101 is Asp, Ala or Met; Xaa at position 105 is Asn or Glu;Xaa at position 109 is Arg, Glu or Leu; Xaa at position 112 is Thr orGln; Xaa at position 116 is Lys, Val, Trp or Ser; Xaa at position 117 isThr or Ser; Xaa at position 120 is Asn, Gln, or His; Xaa at position 123is Ala or Glu; with the proviso that from four to forty-four of theamino acids designated by Xaa are different from the corresponding aminoacids of native human interleukin-3; or a polypeptide havingsubstantially the same structure and substantially the same biologicalactivity.
 14. A polypeptide according to claim 13 wherein Xaa atposition 18 is Ile; Xaa at position 19 is Ala, or Ile; Xaa at position20 is Pro, or Leu; Xaa at position 23 is Ala, or Leu; Xaa at position 25is His; Xaa at position 29 is Arg, Val, or Ile; Xaa at position 32 isAla, Asn or Arg; Xaa at position 34 is Ser; Xaa at position 37 is Pro orSer; Xaa at position 38 is Ala; Xaa at position 42 is Ala, Ser, Asp, orAsn; and Xaa at position 45 is Val or Met; Xaa at position 46 is Ser.15. A polypeptide according to claim 13 wherein Xaa at position 49 isIle, or Leu, or Asp; Xaa at position 50 is Asp; Xaa at position 51 isArg or Ser; Xaa at position 55 is Leu or Thr; Xaa at position 56 is Ser;Xaa at position 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaaat position 62 is Val, or Pro; Xaa at position 63 is His; Xaa atposition 65 is Ser; Xaa at position 67 is Asn, or His, or Gln; and Xaaat position 69 is Glu.
 16. A polypeptide according to claim 13 whereinXaa at position 73 is Gly; Xaa at position 76 is Ala, or Pro; Xaa atposition 79 is Arg, or Ser; Xaa at position 82 is Gln or Val, or Trp;Xaa at position 85 is Val; Xaa at position 87 is Ser, or Tyr; Xaa atposition 88 is Trp; Xaa at position 91 is Pro; Xaa at position 93 isSer; Xaa at position 95 is Thr; Xaa at position 98 is Ile or Thr; Xaa atposition 100 is Arg; Xaa at position 101 is Ala, or Met; and Xaa atposition 105 is Glu.
 17. A polypeptide according to claim 13 wherein Xaaat position 109 is Glu, or Leu; Xaa at position 112 is Gln; Xaa atposition 116 is Val, or Trp, or Ser; Xaa at position 117 is Ser; Xaa atposition 120 is Glu or His; and Xaa at position 123 is Glu.
 18. Apolypeptide according to claim 13 wherein Xaa at position 18 is Ile; Xaaat position 19 is Ala, or Ile; Xaa at position 20 is Pro, or Leu; Xaa atposition 23 is Ala, or Leu; Xaa at position 25 is His; Xaa at position29 is Arg or Val, or Ile; Xaa at position 32 is Ala or Asn, or Arg; Xaaat position 34 is Ser; Xaa at position 37 is Pro or Ser; Xaa at position38 is Ala; Xaa at position 42 is Ala or Ser, Asp or Asn; Xaa at position45 is Val or Met; Xaa at position 46 is Ser; Xaa at position 49 is Ile,or Leu, or Asp; Xaa at position 50 is Asp; Xaa at position 51 is Arg, orSer; Xaa at position 55 is Leu or Thr; Xaa at position 56 is Ser; Xaa atposition 59 is Glu or Leu; Xaa at position 60 is Ala or Ser; Xaa atposition 62 is Val, or Pro; Xaa at position 63 is His; Xaa at position65 is Ser; Xaa at position 67 is Asn, or His, or Gln; and Xaa atposition 69 is Glu.
 19. A polypeptide according to claim 13 wherein Xaaat position 73 is Gly; Xaa at position 76 is Ala, or Pro; Xaa atposition 79 is Arg, or Ser; Xaa at position 82 is Gln or Val, or Trp;Xaa at position 85 is Val; Xaa at position 87 is Ser, or Tyr; Xaa atposition 88 is Trp; Xaa at position 91 is Pro; Xaa at position 93 isSer; Xaa at position 95 is Thr; Xaa at position 98 is Ile or Thr; Xaa atposition 100 is Arg; Xaa at position 101 is Ala, or Met; Xaa at position105 is Glu; Xaa at position 109 is Glu, or Leu; Xaa at position 112 isGln; Xaa at position 116 is Val, or Trp, or Ser; Xaa at position 117 isSer; Xaa at position 120 is Glu or His; and Xaa at position 123 is Glu.20. A polypeptide of the formula             1              5               10 (Met_(m)-Ala_(n))_(p)AsnCys Ser Xaa Xaa Xaa Asp Glu Xaa Ile [SEQ ID NO: 130]                15                  20 Xaa His Leu Lys Xaa Pro Pro XaaPro Xaa Leu Asp Xaa      25                 30                 35 XaaAsn Leu Asn Xaa Gln Asp Xaa Xaa Ile Leu Xaa Gln            40                 45 Xaa Asn Leu Arg Xaa Xaa Asn Leu XaaXaa Phe Xaa Xaa 50                  55                 60 Ala Xaa LysXaa Leu Xaa Asn Ala Ser Xaa Ile Glu Xaa        65                  70                 75 Ile Leu Xaa Asn XaaXaa Pro Cys Xaa Pro Xaa Ala Thr                 80                  85Ala Xaa Pro Xaa Arg Xaa Pro Ile Xaa Ile Xaa Xaa Gly    90                  95                   100 Asp Trp Xaa Glu Phe ArgXaa Lys Leu Xaa Phe Tyr Leu             105                110 Xaa XaaLeu Glu Xaa Ala Gln Xaa Gln Gln

wherein m is 0 or 1; n is 0 or 1; p is 0 or 1; Xaa at position 4 is Asnor Ile; Xaa at position 5 is Met, Ala or Ile: Xaa at position 6 is Ile,Pro or Leu; Xaa at position 9 is Ile, Ala or Leu; Xaa at position 11 isThr or His; Xaa at position 15 is Gln, Arg, Val or Ile; Xaa at position18 is Leu, Ala, Asn or Arg; Xaa at position 20 is Leu or Ser; Xaa atposition 23 is Phe, Pro, or Ser; Xaa at position 24 is Asn or Ala; Xaaat position 28 is Gly, Ala, Ser, Asp or Asn; Xaa at position 31 is Gln,Val, or Met; Xaa at position 32 is Asp or Ser; Xaa at position 35 isMet, Ile or Asp; Xaa at position 36 is Glu or Asp; Xaa at position 37 isAsn, Arg or Ser; Xaa at position 41 is Arg, Leu, or Thr; Xaa at position42 is Pro or Ser; Xaa at position 45 is Glu or Leu; Xaa at position 46is Ala or Ser; Xaa at position 48 is Asn, Val or Pro; Xaa at position 49is Arg or His; Xaa at position 51 is Val or Ser; Xaa at position 53 isSer, Asn, His or Gln; Xaa at position 55 is Gln or Glu; Xaa at position59 is Ala or Gly; Xaa at position 62 is Ser, Ala or Pro; Xaa at position65 is Lys, Arg or Ser; Xaa at position 67 is Leu, Glu, or Val; Xaa atposition 68 is Leu, Glu, Val or Trp; Xaa at position 71 is Leu or Val;Xaa at position 73 is Leu, Ser or Tyr; Xaa at position 74 is Ala or Trp;Xaa at position 77 is Ala or Pro; Xaa at position 79 is Pro or Ser; Xaaat position 81 is His or Thr; Xaa at position 84 is His, Ile, or Thr;Xaa at position 86 is Lys or Arg; Xaa at position 87 is Asp, Ala or Met;Xaa at position 91 is Asn or Glu; Xaa at position 95 is Arg, Glu, Leu;Xaa at position 98 Thr or Gln; Xaa at position 102 is Lys, Val, Trp orSer; Xaa at position 103 is Thr or Ser; Xaa at position 106 is Asn, Gln,or His; Xaa at position 109 is Ala or Glu; with the proviso that fromfour to forty-four of the amino acids designated by Xaa are differentfrom the corresponding amino acids of native (15-125) humaninterleukin-3; or a polypeptide having substantially the same structureand substantially the same biological activity.
 21. A polypeptideaccording to claim 20 wherein Xaa at position 4 is Ile; Xaa at position5 is Ala, or Ile; Xaa at position 6 is Pro, or Leu; Xaa at position 9 isAla, or Leu; Xaa at position 11 is His; Xaa at position 15 is Arg orVal, or Ile; Xaa at position 18 is Ala or Asn, or Arg; Xaa at position20 is Ser; Xaa at position 23 is Pro or Ser; Xaa at position 24 is Ala;Xaa at position 28 is Ala or Ser, or Asp, or Asn; Xaa at position 31 isVal or Met; and Xaa at position 32 is Ser.
 22. A polypeptide accordingto claim 20 wherein Xaa at position 35 is Ile, or Leu, or Asp; Xaa atposition 36 is Asp; Xaa at position 37 is Arg, or Ser; Xaa at position41 is Leu or Thr; Xaa at position 42 is Ser; Xaa at position 45 is Gluor Leu; Xaa at position 46 is Ala or Ser; Xaa at position 48 is Val, orPro; Xaa at position 49 is His; Xaa at position 51 is Ser; Xaa atposition 53 is Asn, or His, or Gln; and Xaa at position 55 is Glu.
 23. Apolypeptide according to claim 20 wherein Xaa at position 59 is Gly; Xaaat position 62 is Ala, or Pro; Xaa at position 65 is Arg, or Ser; Xaa atposition 67 is Gln or Val; Xaa at position 68 is Glu, or Val, or Trp;Xaa at position 71 is Val; Xaa at position 73 is Ser, or Tyr; Xaa atposition 74 is Trp; Xaa at position 77 is Pro; Xaa at position 79 isSer; Xaa at position 81 is Thr; Xaa at position 84 is Ile or Thr; Xaa atposition 86 is Arg; Xaa at position 87 is Ala, or Met; and Xaa atposition 91 is Glu.
 24. A polypeptide according to claim 20 wherein Xaaat position 95 is Glu, or Leu; Xaa at position 98 ia Gln; Xaa atposition 102 is Val, or Trp, or Ser; Xaa at position 103 is Ser; Xaa atposition 106 is Glu or His; and Xaa at position 109 is Glu.
 25. Apolypeptide according to claim 20 wherein Xaa at position 4 is Ile; Xaaat position 5 is Ala, or Ile; Xaa at position 6 is Pro, or Leu; Xaa atposition 9 is Ala, or Leu; Xaa at position 11 is His; Xaa at position 15is Arg or Val, or Ile; Xaa at position 18 is Ala or Asn, or Arg; Xaa atposition 20 is Ser; Xaa at position 23 is Pro or Ser; Xaa at position 24is Ala; Xaa at position 28 is Ala or Ser, or Asp, or Asn; Xaa atposition 31 is Val or Met; Xaa at position 32 is Ser; Xaa at position 35is Ile, or Leu, or Asp; Xaa at position 36 is Asp; Xaa at position 37 isArg, or Ser; Xaa at position 41 is Leu or Thr; Xaa at position 42 isSer; Xaa at position 45 is Glu or Leu; Xaa at position 46 is Ala or Ser;Xaa at position 48 is Val, or Pro; Xaa at position 49 is His; Xaa atposition 51 is Ser; Xaa at position 53 is Asn, or His, or Gln; and Xaaat position 55 is Glu.
 26. A polypeptide according to claim 20 whereinXaa at position 59 is Gly; Xaa at position 62 is Ala, or Pro; Xaa atposition 65 is Arg, or Ser; Xaa at position 67 is Gln or Val; Xaa atposition 68 is Glu, or Val, or Trp; Xaa at position 71 is Val; Xaa atposition 73 is Ser, or Tyr; Xaa at position 74 is Trp; Xaa at position77 is Pro; Xaa at position 79 is Ser; Xaa at position 81 is Thr; Xaa atposition 84 is Ile or Thr; Xaa at position 86 is Arg; Xaa at position 87is Ala, or Met; Xaa at position 91 is Glu; Xaa at position 95 is Glu, orLue; Xaa at position 98 is Gln; Xaa at position 102 is Val, or Trp, orSer; Xaa at position 103 is Ser; Xaa at position 106 is Glu or His; andXaa at position 109 is Glu.
 27. A polypeptide according to claim 20which is selected from      Asn Cys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu [SEQ ID NO:66] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn AsnLeu Asn Ala Gln Asp Val Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro AsnLeu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile GluSer Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro ThrArg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Gln Phe Arg Arg Lys LeuThr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln ;      Asn CysSer Ile Met Ile Asp Gln Ile Ile His His Leu Lys [SEQ ID NO:67] Arg ProPro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Gln Asp Met Asp Ile LeuMet Gln Asn Asn Leu Arg Arg Pro Asn Leu Gln Ala Phe Asn Arg Ala Vai LysSer Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Gln Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Gln AsnAla Gln Ala Gln Gln;      Asn Cys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu Lys [SEQ ID NO:68] Val Pro Pro Ala Pro Leu Leu Asp Ser Asn AsnLeu Asn Ser Gln Asp Met Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro AsnLeu Alu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile GbSer Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro ThrArg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys LeuThr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln;      Asn CysSer Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys [SEQ ID NO:69] Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile LeuMet Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala Phe Val Arg Ala Val LysAsn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu AsnAla Gln Ala Gln Gln;      Asn Cys Ser Asn Met Ile Asp Glu Ile Ile ThrHis Leu Lys [SEQ ID NO:70] Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro AsnLeu Glu Ser Phe Val Arq Ala Val Lys Asn Leu Glu Asn Ala Ser Ala Ile GluSer Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro ThrArg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys LeuThr Phe Tyr Leu Lys Thr Leu Gln Asn Ala Gln Ala Gln Gln;      Asn CysSer Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys [SEQ ID NO:71] Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile LeuMet Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val Arg Ala Val LysHis Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu AsoAla Gln Ala Gln Gln;      Asn Cys Ser Asn Met Ile Asp Gln Ile Ile ThrHis Leu Lys [SEQ ID NO:72] Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Gln Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro AsnLeu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Arg Lys LeuThr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln;      Asn CysSer Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys [SEQ ID NO:73] Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Glu Asp Gln Asp Ile LeuMet Gln Asn Asn Leu Arg Arg Pro Aso Leu Gln Ala Phe Asn Arg Ala Val LysSer Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala GlyAsp Trp Gln Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Gln AsnAla Gln Ala Gln Gln;      Asn Cys Ser Asn Met Ile Asp Gln Ile Ile ThrHis Leu Lys [SEQ ID NO:74] Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Gln Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro AsnLeu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Ala Ile GlnSer Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro ThrArg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Gln Phe Arg Gln Lys LeuThr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln Gln Gln Gln;      Asn CysSer Asn Met Ile Asp Glu Ile Ile Thr His Leu Lys [SEQ ID NO:75] Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln Asp Gln Asp Ile LeuMet Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val LysSer Leu Gln Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Glu Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Ser Leu Gln HisAla Gln Glu Gln Gln;      Asn Cys Ser Asn Met Ile Asp Glu Ile Ile ThrHis Leu Lys [SEQ ID NO:76] Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Gln Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro AsnLeu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GlnAla Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Gln Lys LeuThr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln Gln Gln Gln;      Asn CysSer Asn Met Ile Asp Gln Ile Ile Thr His Leu Lys [SEQ ID NO:77] Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln Asp Gln Asp Ile LeuMet Glu Asn Asn Leu Arg Arg Pro Asn Leu Glu Ala Phe Asn Arg Ala Val LysSer Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala GlyAsp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu GlnAla Gln Glu Gln Gln;      Asn Cys Ser Asn Met Ile Asp Glu Ile Ile ThrHis Leu Lys [SEQ ID NO:78] Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro AsnLeu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys LeuThr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln;      Asn CysSer Ile Met Ile Asp Glu Ile Ile His His Leu Lys [SEQ ID NO:79] Arg ProPro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Glu Asp Val Asp Ile LeuMet Glu Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala Val LysAsn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Gln AsnAla Gln Ala Gln Gln;      Asn Cys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu Lys [SEQ ID NO:80] Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn AsnLeu Asn Ser Gln Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro AsnLeu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala Ile GluSer Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro ThrArg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Gln Phe Arg Arg Lys LeuThr Phe Tyr Leu Lys Thr Leu Gln Asn Ala Gln Ala Gln Gln;      Asn CysSer Ile Met Ile Asp Gln Ile Ile His His Leu Lys [SEQ ID NO:81] Val ProPro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Gln Asp Met Asp Ile LeuMet Gln Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala Phe Val Arg Ala Val LysAsn Leu Gln Asn Ala Ser Ala Ile Gln Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu AsnAla Gln Ala Gln Gln; Met Ala Asn Cys Ser Asn Met Ile Asp Glu Ile Ile ThrHis Leu [SEQ ID NO:82] Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Glu Asp Gln Asp Ile Leu Met Glu Asn Asn Leu Arg Arg Pro AsnLeu Gln Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Gln Phe Arg Gln Lys LeuThr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln Glu Gln Gln; Met Ala Asn CysSer Asn Met Ile Asp Gln Ile Ile Thr His Leu [SEQ ID NO:83] Lys Gln ProPro Leu Pro Leu Leu Asp Phe Asn Asn Leu Asn Gly Gln Asp Gln Asp Ile LeuMet Gln Asn Asn Leu Arg Arg Pro Asn Leu Gln Ala Phe Asn Arg Ala Val LysSer Leu Gln Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala GlyAsp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Gln GlnAla Gln Gln Gln Gln; Met Ala Asn Cys Ser Asn Met Ile Asp Gln Ile Ile ThrHis Leu [SEQ ID NO:84] Lys Gln Pro Pro Leu Pro Leu Leu Asp Phe Asn AsnLeu Asn Gly Glu Asp Gln Asp Ile Leu Met Gln Asn Asn Leu Arg Arg Pro AsnLeu Glu Ala Phe Asn Arg Ala Val Lys Ser Leu Gln Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Gln Phe Arg Gln Lys LeuThr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Gln Gln Gln; Met Ala Asn CysSer Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ ID NO:85] Lys Arg ProPro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Gln Asp Val Asp Ile LeuMet Glu Arg Asn Leu Arg Leu Pro Asn Leu Gln Ser Phe Val Arg Ala Val LysAsn Leu Glu Asn Ala Ser Ala Ile Gln Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Gln Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Gln AsnAla Gln Ala Gln Gln; Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu [SEQ ID NO:86] Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn AsnLeu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro AsnLeu Leu Ala Phe Val Arg Ala Val Lys His Leu Glu Asn Ala Ser Ala Ile GluSer Ile Leu Lys Asn Leu Leu Pro Cys Leu Pro Leu Ala Thr Ala Ala Pro ThrArg His Pro Ile His Ile Lys Asp Gly Asp Trp Asn Glu Phe Arg Arg Lys LeuThr Phe Tyr Leu Lys Thr Leu Glu Asn Ala Gln Ala Gln Gln; Met Ala Asn CysSer Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ ID NO:87] Lys Val ProPro Ala Pro Leu Leu Asp Ser Asn Asn Leu Asn Ser Glu Asp Met Asp Ile LeuMet Glu Arg Asn Leu Arg Leu Pro Asn Leu Leu Ala Phe Val Arg Ala Val LysAsn Leu Glu Asn Ala Ser Ala Ile Glu Ser Ile Leu Lys Asn Leu Leu Pro CysLeu Pro Leu Ala Thr Ala Ala Pro Thr Arg His Pro Ile His Ile Lys Asp GlyAsp Trp Asn Glu Phe Arg Arg Lys Leu Thr Phe Tyr Leu Lys Thr Leu Glu AsnAla Gln Ala Gln Gln; Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile HisHis Leu [SEQ ID NO:88] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn AsnLeu Asn Ala Glu Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro AsnLeu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys LeuThr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln; Met Ala Asn CysSer Ile Met Ile Asp Gln Ile Ile His His Leu [SEQ ID NO:89] Lys Arg ProPro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Gln Asp Met Asp Ile LeuMet Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val Arg Ala Val LysHis Leu Glu Asn Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala GlyAsp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu Gln GlnAla Gln Gln Gln Gln; Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu [SEQ ID NO:90] Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn AsnLeu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro AsnLeu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser Gly Ile GlnAla Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys LeuThr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln Glu Gln Gln; Met Ala Asn CysSer Ile Met Ile Asp Gln Ile Ile His His Leu [SEQ ID NO:91] Lys Arg ProPro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Gln Asp Val Asp Ile LeuMet Gln Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala Val LysAsn Leu Gln Asn Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Val Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala GlyAsp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu GlnAla Gln Glu Gln Gln; Met Ala Asn Gys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu [SEQ ID NO:92] Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn AsnLeu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro AsnLeu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser Gly Ile GlnAla Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Gln Phe Arg Gln Lys LeuThr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Gln Gln Gln; Met Ala Asn CysSer Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ ID NO:93] Lys Arg ProPro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile LeuMet Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val Arg Ala Val LysHis Leu Glu Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg Asn Leu Val Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala GlyAsp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Ser Leu Glu HisAla Gln Glu Gln Gln; Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile HisHis Leu [SEQ ID NO:94] Lys Val Pro Pro Ala Pro Leu Leu Asp Ser Asn AsnLeu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg Asn Leu Arg Leu Pro AsnLeu Leu Ala Phe Val Arg Ala Val Lys Asn Leu Glu Asn Ala Ser Gly Ile GluAla Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro SerArg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys LeuThr Phe Tyr Leu Val Ser Leu Glu His Ala Gln Glu Gln Gln; Met Ala Asn CysSer Ile Met Ile Asp Glu Ile Ile His His Leu Lys Arg Pro Pro Asn Pro LeuLeu Asp Pro Asn Asn Leu Asn Ser Glu Asp Met Asp Ile Leu Met Glu Arg AsnLeu Arg Thr Pro Asn [SEQ ID NO:95] Leu Leu Ala Phe Val Arg Ala Val LysHis Leu Glu Asn Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Val Pro CysLeu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Thr Ile Lys Ala GlyAsp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Gln GlnAla Gln Gln Gln Gln; and Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile IleHis His Leu [SEQ ID NO:96] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro AsnAsn Leu Asn Ala Gln Asp Val Asp Ile Leu Met Glu Arg Asn Leu Arg Leu ProAsn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala Ser Gly IleGlu Ala Ile Leu Arg Asn Leu Val Pro Cys Leu Pro Ser Ala Thr Ala Ala ProSer Arg His Pro Ile Thr Ile Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu LysLeu Thr Phe Tyr Leu Val Ser Leu Gln His Ala Gln Gln Gln Gln. Met Ala AsnCys Ser Ile Met Ile Asp Glu Ile Ile His His Leu [SEQ ID NO.: 296] LysArg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Gln Asp Val AspIle Leu Met Asp Arg Asn Leu Arg Leu Ser Asn Leu Glu Ser Phe Val Arg AlaVal Lys Asn Leu Glu Asn Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu GlnPro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile LysAla Gly Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr LeuGlu Gln Ala Gln Glu Gln Gln. Met Ala Asn Cys Ser Ile Met Ile Asp Glu AlaIle His His Leu [SEQ ID NO.: 300] Lys Arg Pro Pro Ala Pro Ser Leu AspPro Aso Asn Leu Asn Asp Glu Asp Met Ser Ile Leu Met Gln ArgAsn Leu ArgLeu Pro Asn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Gln Asn Ala SerGly Ile Glu Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser Ala Thr AlaAla Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln Glu Phe ArgGln Lys Leu Thr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln Gln Gln Gln MetAla Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu [SEQ ID NO.:301] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Asp Gln AspMet Ser Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn Leu Gln Ser Phe ValArg Ala Val Lys Asn Leu Gln Asn Ala Ser Gly Ile Gln Ala Ile Leu Arg AsnLeu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile IleIle Lys Ala Gly Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu ValThr Leu Glu Gln Ala Gln Gln Gln Gln Met Ala Asn Cys Ser Ile Met Ile AspGlu Ile Ile His His Leu [SEQ ID NO.: 308] Lys Arg Pro Pro Ala Pro LeuLeu Asp Pro Asn Asn Leu Asn Ala Glu Asp Val Asp Ile Leu Met Asp Arg AsnLeu Arg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala Val Lys Asn Leu Glu AsnAla Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro Ser AlaThr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp Gln GluPhe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala Gln Glu GlnGln Met Ala Asn Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu [SEQ IDNO.: 309] Lys Arg Pro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn AspGln Asp Val Ser Ile Leu Met Gln Arg Asn Leu Arg Leu Pro Asn Leu Gln SerPhe Val Arg Ala Val Lys Aso Leu Gln Asn Ala Ser Gly Ile Gln Ala Ile LeuArg Aso Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His ProIle Ile Ile Lys Ala Gly Asp Trp Gln Gln Phe Arg Glu Lys Leu Thr Phe TyrLeu Val Thr Leu Glu Gln Ala Gln Glu Gln Gln Met Ala Asa Cys Ser Ile MetIle Asp Glu Ile Ile His His Leu [SEQ ID NO.: 310] Lys Arg Pro Pro AlaPro Leu Leu Asp Pro Asn Asn Leu Asn Asp Glu Asp Met Ser Ile Leu Met GluArg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala Val Lys Asn LeuGlu Asa Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu ProSer Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp TrpGln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu Val Thr Leu Glu Gln Ala GlnGln Gln Gln Met Ala Tyr Pro Gln Thr Asp Tyr Lys Asp Asp Asp Asp Lys Asn[SEQ ID NO. 315] Cys Ser Ile Met Ile Asp Gln Ile Ile His His Leu Lys ArgPro Pro Ala Pro Leu Leu Asp Pro Asn Asn Leu Asn Ala Gln Asp Val Asp IleLeu Met Gln Arg Asn Leu Arg Leu Pro Asn Leu Glu Ser Phe Val Arg Ala ValLys Asn Leu Glu Asn Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln ProCys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys AlaGly Asp Trp Gln Gln Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu GluGln Ala Gln Gln Gln Gln Met Ala Tyr Pro Gln Thr Asp Tyr Lys Asp Asp AspAsp Lys Asn [SEQ ID NO.: 316] Cys Ser Ile Met Ile Asp Gln Ile Ile HisHis Leu Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser GlnAsp Met Asp Ile Leu Met Gln Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala PheVal Arg Ala Val Lys His Leu Gln Asn Ala Ser Gly Ile Gln Ala Ile Leu ArgAsn Leu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro IleIle Ile Lys Ala Gly Asp Trp Gln Gln Phe Arg Glu Lys Leu Thr Phe Tyr LeuVal Thr Leu Gln Gln Ala Gln Gln Gln Gln Met Ala Asn Cys Ser Ile Met IleAsp Gln Leu Ile His His Leu [SEQ ID NO.: 318] Lys Ile Pro Pro Asn ProSer Leu Asp Ser Ala Asn Leu Asn Ser Gln Asp Val Ser Ile Leu Met Gln ArgAsn Leu Arg Thr Pro Asn Leu Leu Ala Phe Val Arg Ala Val Lys His Leu GlnAsn Ala Ser Gly Ile Gln Ala Ile Leu Arg Asn Leu Gln Pro Cys Leu Pro SerAla Thr Ala Ala Pro Ser Arg His Pro Ile Ile Ile Lys Ala Gly Asp Trp GlnGlu Phe Arg Gln Lys Leu Thr Phe Tyr Leu Val Thr Leu Gln Gln Ala Gln GlnGln Gln.


28. A pharmaceutical composition for the treatment of hematopoietic celldeficiencies comprising a therapeutically effective amount of a mutanthuman interleukin-3 polypeptide selected from the group consisting of apolypeptide of claim 1, a polypeptide of claim 2, a polypeptide of claim3, a polypeptide of claim 4, a polypeptide of claim 5, a polypeptide ofclaim 6, a polypeptide of claim 7, a polypeptide of claim 8, apolypeptide of claim 9, a polypeptide of claim 10, a polypeptide ofclaim 11, a polypeptide of claim 12, a polypeptide of claim 13, apolypeptide of claim 14, a polypeptide of claim 15, a polypeptide ofclaim 16, a polypeptide of claim 17; a polypeptide of claim 18, apolypeptide of claim 19, a polypeptide of claim 20, a polypeptide ofclaim 21, a polypeptide of claim 22, a polypeptide of claim 23, apolypeptide of claim 24, a polypeptide of claim 25, a polypeptide ofclaim 26 and a polypeptide of claim 27, and a pharmaceuticallyacceptable carrier.
 29. A pharmaceutical composition according to claim28 for the treatment of hematopoietic cell deficiencies comprising atherapeutically effective amount of a polypeptide having an amino acidsequence corresponding to SEQ ID NO:88 and a pharmaceutically acceptablecarrier.
 30. A pharmaceutical composition according to claim 28 for thetreatment of hematopoietic cell deficiencies comprising atherapeutically effective amount of a polypeptide having an amino acidsequence corresponding to SEQ ID NO:89 and a pharmaceutically acceptablecarrier.
 31. A pharmaceutical composition according to claim 28 for thetreatment of hematopoietic cell deficiencies comprising atherapeutically effective amount of a polypeptide having an amino acidsequence corresponding to SEQ ID NO:90 and a pharmaceutically acceptablecarrier.
 32. A pharmaceutical composition according to claim 28 for thetreatment of hematopoietic cell deficiencies comprising atherapeutically effective amount of a polypeptide selected from thegroup consisting of a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:66; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:67; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:68; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:69; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:70; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:71; apolypeptide having an amino acid sequence corresponding to SEQ ID NO:72;a polypeptide having an amino acid sequence corresponding to SEQ IDNO:73; a polypeptide having an amino acid sequence corresponding to SEQID NO:74; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:75; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:76; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:77; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:78; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:79; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:80; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:81; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:82; apolypeptide having an amino acid sequence corresponding to SEQ ID NO:83;a polypeptide having an amino acid sequence corresponding to SEQ IDNO:84; a polypeptide having an amino acid sequence corresponding to SEQID NO:85; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:86; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:87; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:91; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:92; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:93; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:94; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:95; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:96; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:258; a polypeptide having an amino acid sequence corresponding to SEQID NO:259; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:260; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:261; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:262; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:263; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:278; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:279; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:314; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:315; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:316; a polypeptide having an amino acid sequence corresponding to SEQID NO:264; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:265; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:266; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:267; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:268; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:269; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:270; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:271; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:272; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:273; a polypeptide having an amino acid sequence corresponding to SEQID NO:274; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:275; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:276; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:277; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:280; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:281; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:282; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:283; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:284; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:285; a polypeptide having an amino acid sequence corresponding to SEQID NO:286; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:287; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:288; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:289; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:299; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:300; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:301; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:302; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:303; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:304; a polypeptide having an amino acid sequence corresponding to SEQID NO:305; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:306; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:307; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:308; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:309; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:310; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:311; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:312; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:313; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:314; a polypeptide having an amino acid sequence corresponding to SEQID NO:317; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:318; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:319; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:320; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:321; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:322; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:323; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:324; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:325; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:326; and a pharmaceutically acceptable carrier.
 33. A method ofstimulating the production of hematopoietic cells which comprisesadministering a therapeutically effective amount of a mutant humaninterleukin-3 polypeptide selected from the group consisting of apolypeptide of claim 1, a polypeptide of claim 2, a polypeptide of claim3, a polypeptide of claim 4, a polypeptide of claim 5, a polypeptide ofclaim 6, a polypeptide of claim 7, a polypeptide of claim 8, apolypeptide of claim 9, a polypeptide of claim 10, a polypeptide ofclaim 11, a polypeptide of claim 12, a polypeptide of claim 13, apolypeptide of claim 14, a polypeptide of claim 15, a polypeptide ofclaim 16, a polypeptide of claim 17; a polypeptide of claim 18, apolypeptide of claim 19, a polypeptide of claim 20, a polypeptide ofclaim 21, a polypeptide of claim 22, a polypeptide of claim 23, apolypeptide of claim 24, a polypeptide of claim 25, a polypeptide ofclaim 26, a polypeptide of claim 27, to a patient in need of suchtreatment.
 34. A method according to claim 33 of stimulating theproduction of hematopoietic cells which comprises administering atherapeutically effective amount of a polypeptide having an amino acidsequence corresponding to SEQ ID NO:88.
 35. A method according to claim33 of stimulating the production of hematopoietic cells which comprisesadministering a therapeutically effective amount of a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:89.
 36. A methodaccording to claim 33 of stimulating the production of hematopoieticcells which comprises administering a therapeutically effective amountof a polypeptide having an amino acid sequence corresponding to SEQ IDNO:90.
 37. A method according to claim 33 of stimulating the productionof hematopoietic cells which comprises administering a therapeuticallyeffective amount of a polypeptide selected from the group consisting ofa polypeptide having an amino acid sequence corresponding to SEQ IDNO:66; a polypeptide having an amino acid sequence corresponding to SEQID NO:67; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:68; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:69; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:70; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:71; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:72; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:73; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:74; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:75; apolypeptide having an amino acid sequence corresponding to SEQ ID NO:76;a polypeptide having an amino acid sequence corresponding to SEQ IDNO:77; a polypeptide having an amino acid sequence corresponding to SEQID NO:78; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:79; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:80; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:81; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:82; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:83; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:84; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:85; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:86; apolypeptide having an amino acid sequence corresponding to SEQ ID NO:87;a polypeptide having an amino acid sequence corresponding to SEQ IDNO:91; a polypeptide having an amino acid sequence corresponding to SEQID NO:92; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:93; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:94; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:95; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:96; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:258; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:259; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:260; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:261; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:262; a polypeptide having an amino acid sequence corresponding to SEQID NO:263; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:278; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:279; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:314; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:315; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:316; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:264; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:265; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:266; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:267; a polypeptide having an amino acid sequence corresponding to SEQID NO:268; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:269; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:270; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:271; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:272; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:273; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:274; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:275; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:276; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:277; a polypeptide having an amino acid sequence corresponding to SEQID NO:280; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:281; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:282; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:283; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:284; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:285; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:286; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:287; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:288; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:289; a polypeptide having an amino acid sequence corresponding to SEQID NO:299; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:300; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:301; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:302; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:303; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:304; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:305; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:306; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:307; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:308; a polypeptide having an amino acid sequence corresponding to SEQID NO:309; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:310; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:311; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:312; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:313; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:314; a polypeptide having anamino acid sequence corresponding to SEQ ID NO:317; a polypeptide havingan amino acid sequence corresponding to SEQ ID NO:318; a polypeptidehaving an amino acid sequence corresponding to SEQ ID NO:319; apolypeptide having an amino acid sequence corresponding to SEQ IDNO:320; a polypeptide having an amino acid sequence corresponding to SEQID NO:321; a polypeptide having an amino acid sequence corresponding toSEQ ID NO:322; a polypeptide having an amino acid sequence correspondingto SEQ ID NO:323; a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:324; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:325; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:326; to a patient in need ofsuch treatment.
 38. A recombinant DNA sequence comprising vector DNA anda DNA that encodes a polypeptide selected from the group consisting of apolypeptide of claim 1, a polypeptide of claim 2, a polypeptide of claim3, a polypeptide of claim 4, a polypeptide of claim 5, a polypeptide ofclaim 6, a polypeptide of claim 7, a polypeptide of claim 8, apolypeptide of claim 9, a polypeptide of claim 10, a polypeptide ofclaim 11, a polypeptide of claim 12, a polypeptide of claim 13, apolypeptide of claim 14, a polypeptide of claim 15, a polypeptide ofclaim 16, a polypeptide of claim 17; a polypeptide of claim 18, apolypeptide of claim 19, a polypeptide of claim 20, a polypeptide ofclaim 21, a polypeptide of claim 22, a polypeptide of claim 23, apolypeptide of claim 24, a polypeptide of claim 25, a polypeptide ofclaim 26, or a polypeptide of claim
 27. 39. A recombinant DNA sequenceaccording to claim 38 comprising vector DNA and a DNA having anucleotide sequence corresponding to SEQ ID NO:97.
 40. A recombinant DNAsequence according to claim 38 comprising vector DNA and a DNA having anucleotide sequence corresponding to SEQ ID NO:100 or
 103. 41. Arecombinant DNA sequence according to claim 38 comprising vector DNA anda DNA having a nucleotide sequence corresponding to SEQ ID NO:161.
 42. Arecombinant DNA sequence according to claim 38 comprising vector DNA anda DNA selected from a DNA having a nucleotide sequence corresponding toSEQ ID NO:98; a DNA having a nucleotide sequence corresponding to SEQ IDNO:99; a DNA having a nucleotide sequence corresponding to SEQ IDNO:101; a DNA having a nucleotide sequence corresponding to SEQ IDNO:102; a DNA having a nucleotide sequence corresponding to SEQ IDNO:104; a DNA having a nucleotide sequence corresponding to SEQ IDNO:105; a DNA having a nucleotide sequence corresponding to SEQ IDNO:106; a DNA having a nucleotide sequence corresponding to SEQ IDNO:107; a DNA having a nucleotide sequence corresponding to SEQ IDNO:108; a DNA having a nucleotide sequence corresponding to SEQ IDNO:109; a DNA having a nucleotide sequence corresponding to SEQ IDNO:110; a DNA having a nucleotide sequence corresponding to SEQ IDNO:111; a DNA having a nucleotide sequence corresponding to SEQ IDNO:112; a DNA having a nucleotide sequence corresponding to SEQ IDNO:113; a DNA having a nucleotide sequence corresponding to SEQ IDNO:114; a DNA having a nucleotide sequence corresponding to SEQ ID NO:115; a DNA having a nucleotide sequence corresponding to SEQ ID NO:116;a DNA having a nucleotide sequence corresponding to SEQ ID NO:117; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:118; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:119; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:120; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:121; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:122; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:123; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:124; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:125; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:126; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:127; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:160; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:161; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:398; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:399; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:346; a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:347 a DNA havinga nucleotide sequence corresponding to SEQ ID NO:303 a DNA having anucleotide sequence corresponding to SEQ ID NO:404 a DNA having anucleotide sequence corresponding to SEQ ID NO:405 a DNA having anucleotide sequence corresponding to SEQ ID NO:332 a DNA having anucleotide sequence corresponding to SEQ ID NO:333 a DNA having anucleotide sequence corresponding to SEQ ID NO:334 a DNA having anucleotide sequence corresponding to SEQ ID NO:335 a DNA having anucleotide sequence corresponding to SEQ ID NO:336 a DNA having anucleotide sequence corresponding to SEQ ID NO:337 a DNA having anucleotide sequence corresponding to SEQ ID NO:338 a DNA having anucleotide sequence corresponding to SEQ ID NO:339 a DNA having anucleotide sequence corresponding to SEQ ID NO:340 a DNA having anucleotide sequence corresponding to SEQ ID NO:341 a DNA having anucleotide sequence corresponding to SEQ ID NO:342 a DNA having anucleotide sequence corresponding to SEQ ID NO:343 a DNA having anucleotide sequence corresponding to SEQ ID NO:344 a DNA having anucleotide sequence corresponding to SEQ ID NO:345 a DNA having anucleotide sequence corresponding to SEQ ID NO:348 a DNA having anucleotide sequence corresponding to SEQ ID NO:349 a DNA having anucleotide sequence corresponding to SEQ ID NO:350 a DNA having anucleotide sequence corresponding to SEQ ID NO:352 a DNA having anucleotide sequence corresponding to SEQ ID NO:353 a DNA having anucleotide sequence corresponding to SEQ ID NO:354 a DNA having anucleotide sequence corresponding to SEQ ID NO:355 a DNA having anucleotide sequence corresponding to SEQ ID NO:356 a DNA having anucleotide sequence corresponding to SEQ ID NO:357 a DNA having anucleotide sequence corresponding to SEQ ID NO:358 a DNA having anucleotide sequence corresponding to SEQ ID NO:359 a DNA having anucleotide sequence corresponding to SEQ ID NO:360 a DNA having anucleotide sequence corresponding to SEQ ID NO:361 a DNA having anucleotide sequence corresponding to SEQ ID NO:362 a DNA having anucleotide sequence corresponding to SEQ ID NO:363 a DNA having anucleotide sequence corresponding to SEQ ID NO:364 a DNA having anucleotide sequence corresponding to SEQ ID NO:365 a DNA having anucleotide sequence corresponding to SEQ ID NO:366 a DNA having anucleotide sequence corresponding to SEQ ID NO:367 a DNA having anucleotide sequence corresponding to SEQ ID NO:368 a DNA having anucleotide sequence corresponding to SEQ ID NO:369 a DNA having anucleotide sequence corresponding to SEQ ID NO:370 a DNA having anucleotide sequence corresponding to SEQ ID NO:371 a DNA having anucleotide sequence corresponding to SEQ ID NO:372 a DNA having anucleotide sequence corresponding to SEQ ID NO:373 a DNA having anucleotide sequence corresponding to SEQ ID NO:374 a DNA having anucleotide sequence corresponding to SEQ ID NO:375 a DNA having anucleotide sequence corresponding to SEQ ID NO:376 a DNA having anucleotide sequence corresponding to SEQ ID NO:377 a DNA having anucleotide sequence corresponding to SEQ ID NO:378 a DNA having anucleotide sequence corresponding to SEQ ID NO:379 a DNA having anucleotide sequence corresponding to SEQ ID NO:380 a DNA having anucleotide sequence corresponding to SEQ ID NO:381 a DNA having anucleotide sequence corresponding to SEQ ID NO:382 a DNA having anucleotide sequence corresponding to SEQ ID NO:384 a DNA having anucleotide sequence corresponding to SEQ ID NO:385 a DNA having anucleotide sequence corresponding to SEQ ID NO:386 a DNA having anucleotide sequence corresponding to SEQ ID NO:387 a DNA having anucleotide sequence corresponding to SEQ ID NO:388 a DNA having anucleotide sequence corresponding to SEQ ID NO:389 a DNA having anucleotide sequence corresponding to SEQ ID NO:390 a DNA having anucleotide sequence corresponding to SEQ ID NO:391 a DNA having anucleotide sequence corresponding to SEQ ID NO:392
 43. A host cellcontaining a recombinant DNA sequence of claim 38 and capable ofexpressing the encoded polypeptide.
 44. A host cell of claim 43containing a recombinant DNA vector comprising vector DNA and a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:97 and capableof expressing the encoded polypeptide.
 45. A host cell of claim 43containing a recombinant DNA vector comprising vector DNA and a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:100 or 103 andcapable of expressing the encoded polypeptide.
 46. A host cell of claim43 containing a recombinant DNA vector comprising vector DNA and a DNAhaving a nucleotide sequence corresponding to SEQ ID NO:161 and capableof expressing the encoded polypeptide.
 47. A method of producing amutant human interleukin-3 polypeptide comprising the steps of: (a)culturing a host cell containing a recombinant DNA sequence comprisingvector DNA and a DNA sequence of claim 38 and capable of expressing theencoded polypeptide under conditions permitting expression of therecombinant DNA; and (b) harvesting the polypeptide from the culture.48. A method according to claim 47 of producing a mutant humaninterleukin-3 polypeptide comprising the steps of: (a) culturing a hostcell containing a recombinant DNA sequence comprising vector DNA and aDNA having a nucleotide sequence corresponding to SEQ ID NO:97 andcapable of expressing the encoded polypeptide under conditionspermitting expression of the recombinant DNA; and (b) harvesting thepolypeptide from the culture.
 49. A method according to claim 47 ofproducing a mutant human interleukin-3 polypeptide comprising the stepsof: (a) culturing a host cell containing a recombinant DNA sequencecomprising vector DNA and a DNA having a nucleotide sequencecorresponding to SEQ ID NO:100 or 103 and capable of expressing theencoded polypeptide under conditions permitting expression of therecombinant DNA; and (b) harvesting the polypeptide from the culture.50. A method according to claim 47 of producing a mutant humaninterleukin-3 polypeptide comprising the steps of: (a) culturing a hostcell containing a recombinant DNA sequence comprising vector DNA and aDNA having a nucleotide sequence corresponding to SEQ ID NO:161 andcapable of expressing the encoded polypeptide under conditionspermitting expression of the recombinant DNA; and (b) harvesting thepolypeptide from the culture.
 51. A vector containing a gene having aDNA sequence selected from the group consisting of: a DNA having anucleotide sequence corresponding to SEQ ID NO:97; a DNA having anucleotide sequence corresponding to SEQ ID NO:100; a DNA having anucleotide sequence corresponding to SEQ ID NO:103; a DNA having anucleotide sequence corresponding to SEQ ID NO:160; a DNA having anucleotide sequence corresponding to SEQ ID NO:161; a DNA having anucleotide sequence corresponding to SEQ ID NO:404; a DNA having anucleotide sequence corresponding to SEQ ID NO:405; a DNA having anucleotide sequence corresponding to SEQ ID NO:364; a DNA having anucleotide sequence corresponding to SEQ ID NO:368; a DNA having anucleotide sequence corresponding to SEQ ID NO:369; a DNA having anucleotide sequence corresponding to SEQ ID NO:376; a DNA having anucleotide sequence corresponding to SEQ ID NO:377; a DNA having anucleotide sequence corresponding to SEQ ID NO:378; a DNA having anucleotide sequence corresponding to SEQ ID NO:385;
 52. A recombinantDNA vector comprising a promoter, a ribosome binding site, and a signalpeptide directly linked to a DNA sequence encoding a polypeptideselected from the group consisting of a polypeptide having an amino acidsequence corresponding to SEQ ID NO:88; a polypeptide having an aminoacid sequence corresponding to SEQ ID NO:89; and a polypeptide having anamino acid sequence corresponding to SEQ ID NO:90; said vector beingcapable of directing expression of said mutant human interleukin-3polypeptide.
 53. A recombinant DNA vector according to claim 51 whereinthe promoter is AraBAD.
 54. A recombinant DNA vector according to claim51 wherein the ribosome binding site is g10-L.
 55. A recombinant DNAvector according to claim 51 wherein the signal peptide is a lamB signalpeptide.
 56. A recombinant DNA vector according to claim 51 wherein thesignal peptide is the lamB signal peptide depicted in FIG.
 8. 57. Arecombinant DNA vector according to claim 51 wherein the promoter isAraBAD and the ribosome binding site is g10-L.
 58. A recombinant DNAvector according to claim 51 wherein the promoter is AraBAD, theribosome binding site is g10-L, and the signal peptide is a lamB signalpeptide.
 59. A recombinant DNA vector according to claim 51 wherein thepromoter is AraBAD, the ribosome binding site is g10-L, and the signalpeptide is the lamB signal peptide depicted in FIG.
 8. 60. A recombinantbacterial host which comprises the vector of claim 51 wherein said hostsecretes a mutant human interleukin-3 polypeptide selected from thegroup consisting of a polypeptide having an amino acid sequencecorresponding to SEQ ID NO:88; a polypeptide having an amino acidsequence corresponding to SEQ ID NO:89; and a polypeptide having anamino acid sequence corresponding to SEQ ID NO:90.
 61. A polypeptide ofthe formula          1            5                      10 [SEQ IDNO:129] (Met)_(m)-Ala Pro Met Thr Gln Thr Thr Ser Leu Lys Thr             15                   20 Ser Trp Val Asn Cys Ser Xaa Met IleAsp Glu Ile Ile 25                  30             35 Xaa His Leu LysXaa Pro Pro Xaa Pro Leu Leu Asp Xaa        40                  45                  50 Asn Asn Leu Asn XaaGlu Asp Xaa Asp Ile Leu Met Glu                 55                60 XaaAsn Leu Arg Xaa Pro Asn Leu Xaa Xaa Phe Xaa Arg     65                 70                  75 Ala Val Lys Xaa Leu XaaAsn Ala Ser Xaa Ile Glu Xaa             80                  85 Ile LeuXaa Asn Leu Xaa Pro Cys Leu Pro Xaa Ala Thr90                  95               100 Ala Ala Pro Xaa Arg His Pro IleXaa Ile Lys Xaa Gly         105               110               115 AspTrp Xaa Glu Phe Arg Xaa Lys Leu Thr Phe Tyr Leu               120                125 Xaa Thr Leu Glu Xaa Ala Gln XaaGln Gln Thr Thr Leu     130 Ser Leu Ala Ile Phe

wherein m is 0 or 1; Xaa at position 18 is Asn or Ile; Xaa at position25 is Thr or His; Xaa at position 29 is Gln, Arg, or Val; Xaa atposition 32 is Leu, Ala, or Asn; Xaa at position 37 is Phe, Pro, or Ser;Xaa at position 42 is Glu, Ala, or Ser; Xaa at position 45 is Gln, Val,or Met; Xaa at position 51 is Asn or Arg; Xaa at position 55 is Arg,Leu, or Thr; Xaa at position 59 is Glu or Leu; Xaa at position 60 is Alaor Ser; Xaa at position 62 is Asn or Val; Xaa at position 67 is Ser,Asn, or His; Xaa at position 69 is Gln or Glu; Xaa at position 73 is Alaor Gly; Xaa at position 76 is Ser or Ala; Xaa at position 79 is Lys orArg; Xaa at position 82 is Leu, Glu, or Val; Xaa at position 87 is Leuor Ser; Xaa at position 93 is Pro or Ser; Xaa at position 98 is His,Ile, or Thr; Xaa at position 101 is Asp or Ala; Xaa at position 105 isAsn or Glu; Xaa at position 109 is Arg or Glu; Xaa at position 116 isLys or Val; Xaa at position 120 is Asn, Gln, or His; Xaa at position 123is Ala or Glu; with the proviso that from four to twenty-seven of theamino acids designated by Xaa are different from the corresponding aminoacids of native human interleukin-3 and wherein from 1 to 14 of aminoacids 1 to 14 has been deleted from the N-terminus and/or from 1 to 15of amino acids 119 to 133 has been deleted from the C-terminus of thepolypeptide; or a polypeptide having substantially the same structureand substantially the same biological activity.
 62. A method accordingto claim 47 of producing a mutant human interleukin-3 polypeptidecomprising the steps of: (a) culturing a host cell containing arecombinant DNA sequence comprising vector DNA and a DNA having anucleotide sequence corresponding to SEQ ID NO:160 and capable ofexpressing the encoded polypeptide under conditions permittingexpression of the recombinant DNA; and (b) harvesting the polypeptidefrom the culture.
 63. A method according to claim 47 of producing amutant human interleukin-3 polypeptide comprising the steps of: (a)culturing a host cell containing a recombinant DNA sequence comprisingvector DNA and a DNA having a nucleotide sequence corresponding to SEQID NO:161 and capable of expressing the encoded polypeptide underconditions permitting expression of the recombinant DNA; and (b)harvesting the polypeptide from the culture.
 64. A host cell containinga recombinant DNA vector comprising vector DNA and a DNA sequenceselected from the group consisting of: a DNA having a nucleotidesequence corresponding to SEQ ID NO:160; and a DNA having a nucleotidesequence corresponding to SEQ ID NO:161; and capable of expressing theencoded polypeptide.
 65. A polypeptide according to claim 27 which is:[SEQ ID NO:89] Met Ala Asn Cys Ser Ile Met Ile Asp Glu Ile Ile His HisLeu Lys Arg Pro Pro Asn Pro Leu Leu Asp Pro Asn Asn Leu Asn Ser Glu AspMet Asp Ile Leu Met Glu Arg Asn Leu Arg Thr Pro Asn Leu Leu Ala Phe ValArg Ala Val Lys His Leu Glu Asn Ala Ser Gly Ile Glu Ala Ile Leu Arg AsnLeu Gln Pro Cys Leu Pro Ser Ala Thr Ala Ala Pro Ser Arg His Pro Ile IleIle Lys Ala Gly Asp Trp Gln Glu Phe Arg Glu Lys Leu Thr Phe Tyr Leu ValThr Leu Glu Gln Ala Gln Glu Gln Gln.

INTERLEUKIN-3 (IL-3) MULTIPLE MUTATION POLYPEPTIDES